by Dr Huub H. van Rossum Recently, significant improvements have been made in understanding and applying moving average quality control (MA QC) that enable its practical implementation. These include the description of new and laboratory-specific MA QC optimization and validation methods, the online availability thereof, insights into operational requirements, and demonstration of practical implementation. Introduction Moving average quality control (MA QC) is the process of algorithmically averaging obtained test results and using that average for (analytical) quality control purposes. MA QC is generally referred to as patient-based real-time quality control (PBRTQC) because it is one of various methods (e.g. limit checks, delta checks, etc) that use patient results for (real-time) quality control. MA QC was first described over half a century ago as ‘average of normals’ [1]. Since then, it has evolved into a more general MA QC concept not necessarily based on using mean calculations of the obtained ‘normal’ test results [2]. Although MA QC has been available for a few decades, its adoption by laboratories has been limited due to the complexity of setting up the necessary procedures, operational challenges and a lack of evidence to justify its application and demonstrate its value. During the past 5|years, however, significant improvements have been made in the field of MA QC, and research studies have addressed all these issues. Consequently, true practical application of validated MA QC procedures to support analytical quality control in medical laboratories is now possible. Furthermore, the recent improvements may well change the way we perform daily analytical quality control in medical laboratories in the near future. MA QC optimization and validation The recent significant improvements in the field of MA QC include, first and foremost, the description of new methods to design and optimize laboratory-specific MA QC procedures and to enable validation of their actual error-detection performance [2–5]. These methods use realistic MA QC simulations based on laboratory-specific datasets and thus provide objective insights into MA QC error detection [2]. To enable practical implementation, the requirement that the number of MA QC alarms must be manageable is now acknowledged as essential and has been fulfilled when setting up MA QC [2, 6]. The newly developed methods use a novel metric to determine the error-detection performance: that is, the mean or median number of test results needed for error detection. One of the new methods presents these simulation results in bias-detection curves so that the optimal MA QC procedure can be selected, based on its overall error-detection performance [5]. An example of a bias-detection curve and its application is presented in Figure 1. After selecting the optimal MA QC settings, an MA validation chart can be used to obtain objective insights into the overall error-detection performance and the uncertainty thereof. Therefore, this chart can be seen as a validation of the MA QC procedure. An example of an MA validation chart is presented in Figure 2 and shows that the MA QC procedure will almost always (with 97.5% probability) detect a systematic error of −4% (or larger negative errors) within 20 test results.
Importantly, this method has become available to laboratories via the online MA Generator application, enabling them to design their own optimized and validated MA QC procedures [7]. Laboratories can now upload their own datasets of historical results, study potential MA QC settings using this simulation analysis and obtain their own laboratory-specific MA QC settings and MA validation charts. Several laboratories have demonstrated that this tool has enabled them to obtain relevant MA QC settings and thus implement MA QC [8, 9]. Integration of MA QC with internal QC Measurement of internal quality control (iQC) samples is still the cornerstone of analytical quality control as performed in medical laboratories. For many tests, iQC alone is sufficient to assure and control the quality of obtained test results. For some tests, however, iQC itself is insufficient. The reasons for this are related to certain fundamental characteristics of iQC that include: lack of available (stable) control materials, its scheduled character, the risk of using non-commutable control samples and tests with a sigma metric score of ≤4. For several reasons, PBRTQC or, more specifically, MA QC is a particularly valuable and powerful way to support quality assurance in such cases.
First, if no (stable) QC materials are available it is impossible, or it becomes complicated, to use iQC. This is, for example, relevant for the erythrocyte sedimentation rate, serum indices or hemocytometry tests including erythrocyte mean corpuscular volume in particular. MA QC is possible as long as patient results are available. Second, the scheduled character of iQC becomes a limitation and a risk when temporary assay failures or rapid onset of critical errors occur between scheduled iQC. Because a new MA QC value can be calculated for each newly obtained test result, MA QC can be designed as a continuous and real-time QC tool. In this context, detection of temporary assay failure by MA QC between scheduled iQC has been demonstrated for a sodium case [10], and several examples of MA QC detection of rapid onset of critical errors have been published for both chemistry and hematological tests [11]. Third, because PBRTQC methods such as MA QC use obtained patient results, by design there is no commutability issue. Fourth, and finally, for some tests iQC is intrinsically limited in its ability to detect relevant clinical errors, due to the low ratio of biological variations to analytical variations, as reflected in low sigma metric values. Such tests require frequent iQC analysis and application of stringent control rules. However, even with such an intensive and strict iQC set-up, the probability of detecting clinically relevant errors remains limited [12]. In contrast, MA QC has the best error-detection performance for tests with a low sigma value [13].
For all these reasons, MA QC is ideal for supplementing analytical quality control by iQC. Recently, an approach was presented that integrated MA QC into the QC plan when iQC was found to be insufficient [9]. This approach was based on first determining whether one of the abovementioned iQC limitations applied to a test. If so, then iQC alone was considered insufficient and MA QC was studied, using the online MA Generator tool (www.huvaros.com) to obtain optimal MA QC settings and MA QC procedures to support the analytical quality control [7, 9]. The MA QC error-detection performance was validated using MA validation charts. These latter insights into MA QC error detection also enabled iQC measurements to be reduced. The MA QC procedures alone provided significant error-detection performance, so running iQC measures multiple times a day would add only limited error-detection performance. Therefore, it was decided to run the iQC only once a day and add the obtained MA QC procedures to the QC plan.
Others have taken this a step further and studied MA QC not only for tests with limited iQC performance but also for a much larger test selection, in order to reduce the number of iQC measures and more efficiently schedule and apply iQC [4]. This approach has been shown to be successful for a large commercial laboratory with high production numbers. Since the MA QC error-detection performance improves with an increasing number of test results and benefits from a small number of pathological test results, this approach may be particularly valuable to the larger commercial laboratories. For such an approach, the key is objective insights into the error-detection performance of MA QC procedures such as obtained using MA validation charts. Implementation and application of MA QC for real-time QC in medical laboratories The final aspect in which there have been significant improvements in recent years relates to the practical application of MA QC in medical laboratories. Recently, an International Federation of Clinical Chemistry and Laboratory Medicine working group was founded that summarized medical laboratories’ experiences of practically applying MA QC and formulated several recommendations for both MA QC software suppliers and medical laboratories that are working on, or are interested in, implementation of MA QC [14, 15]. Also, a step-by-step roadmap has recently been published to enable MA QC implementation [9]. The first two steps of this roadmap – i.e. selection of tests and obtaining MA QC settings for them – were discussed in the previous two paragraphs.
The next step would be to set up and configure the software used to implement MA QC in medical laboratories. If you are interested in applying MA QC in your laboratory, it is important to review the available software (e.g. analyser, middleware, LIS, third party) and to decide which will be used to run and apply MA QC. Your decision depends not only on the availability of suitable software in or for the laboratory, but also on the actual MA QC functionality present in the software packages.
The minimum software features that are necessary to enable practical implementation have been formulated [2, 15]. In my view, key elements would be that the software supports: exclusion of specified samples (non-patient materials, QC results, extreme results, etc), calculation of relevant MA QC algorithms, applying SD-based as well as non-statistical control limits (plain lower and upper control limits), proper real-time alarming and – depending on the MA QC optimization method – presentation of MA QC in a Levey–Jennings or accuracy graph. Figure 3 presents an example of MA QC in an accuracy graph as operated for real-time QC in my laboratory. To enable effective implementation of MA QC, all of these software features should be configured.
The final implementation step I wish to address here is the design of laboratory protocols for working up MA QC alarms, which determines the extent to which an error detected by an MA QC alarm is acknowledged. An important requirement is that all MA QC alarms should be worked up by means of this protocol.
As previously indicated, because MA QC can generate many more QC results and alarms than iQC, a critical requirement of every MA QC procedure is a manageable number of alarms. As a result, when an MA QC alarm occurs there is a reasonable chance of detecting error.
A first common action as part of the MA QC alarm protocol would be to run iQC. This provides a quick insight into the size of the error and enables rapid confirmation of large errors. As a second step, re-running of recently analysed samples (in addition to running iQC) enables temporary assay failures to be detected and can confirm or exclude errors not necessarily detectable by iQC. Also, finally, a review of recently analysed test results to identify a pre-analytical cause or a single patient with extreme but valid test results is often very useful as part of the MA QC alarm protocol. All these aspects have recently been discussed in greater detail [10, 14]. Conclusions Altogether, the recent developments in the field of PBRTQC and, more specifically, MA QC now – finally – enable true practical implementation of MA QC in medical laboratories and allow more effective and efficient QC plans to be designed. The authors Huub H. van Rossum1,2 PhD 1 Department of Laboratory Medicine, The Netherlands Cancer Institute, Amsterdam, The Netherlands 2 Huvaros, Amsterdam, The Netherlands
E-mail: h.v.rossum@nki.nl
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by Prof. Michael Vogeser, Dr Judy Stone and Prof. Alan Rockwood While analytical standardization and metrological traceability are well-defined terms, ‘methodological standardization’ in clinical mass spectrometry is still in a developing stage. We propose a framework that facilitates the widespread implementation of this highly complex and very powerful technology and is based on two pillars – standardization of the description of LC-MS/MS methods and standardization of the release of clinical test results as a three-step sequence of method validation, batch validation and validation of individual measurements. Mass spectrometry in the clinical laboratory Mass spectrometry (MS)-based methods now play an important role in many clinical laboratories worldwide. To date, areas of application have focused especially on screening for hereditary metabolic diseases, therapeutic drug monitoring, clinical toxicology and endocrinology. In fact, these techniques offer significant advantages over immunoassays and photometry as basic standard technologies in clinical chemistry: high analytical selectivity through true molecular detection; wide range of applications without the need for specific molecular features (as in UV detection or specific epitopes); high multiplexing capacity and information-rich detection; and, in many cases, matrixindependent analyses, thanks to the principle of isotope dilution [1].
Various MS technologies – in particular tandem MS (MS/MS-coupling with molecular fragmentation), time-of-flight (TOF) MS and Orbitrap-MS – with front-end fractionation technologies such as HPLC or UPLC potentially allow very reliable analysis, but the technology itself is no guarantee of this: these techniques have a very high complexity and a wide range of potential sources of error [2] which require comprehensive quality assurance [3–5]. Indeed, the high degree of complexity is still the main hurdle for the application of MS in the special environment of clinical laboratories. Specific challenges of this type of laboratory – in contrast to research and development laboratories – include: heterogeneous mix of staff qualifications; requirement for maximum handling safety when operating a large number of analysis platforms; work around the clock; and direct impact on the outcome of the individual patient.
Indeed, after more than two decades of commercial availability of LC-MS/MS instruments, their application in a global perspective has remained very limited. The translation of MS into fully automated ‘black box’ instruments is underway, but still far from being realized on a large scale [6], with laboratory developed tests (LDTs) still dominating the field of clinical MS applications. Kit solutions for specific analytes provided by the in vitro diagnostics (IVD) industry are becoming increasingly available, but their application also requires a very high level of skills and competence from laboratories.
Two main differences of MS-based LDTs as opposed to standard ‘plug-and-play’ analysis systems in today’s clinical laboratories can be identified: first, the high heterogeneity of device configurations and second, the handling of large amounts of data, from sample list structures to technical metadata analysis.
In fact, the random access working mode is now so widespread in all clinical laboratories that the ‘analytical batch’ is no longer standard in laboratories. In the same way, modern analytical instruments no longer challenge the end users with extensive metadata (such as reaction kinetics or calibration diagrams). To achieve the goal of making the extraordinary and disruptive analytical power of MS fully usable for medicine to an appropriate extent, approaches to master the heterogeneity of platform configurations and to regulate the handling of batches and metadata are urgently needed – and standardization efforts seem to be crucial in this context. Standardization of the method description IVD companies manufacture many different instrument platforms, but each of these platforms is very homogeneous worldwide and is produced in large quantities for years. In contrast, MS platforms in clinical laboratories have to be individually assembled from a very large number of components from many manufacturers (sample preparation modules, autosamplers, high performance pumps, switching valves, chromatography columns, ion sources, mass analysers, vacuum systems, software packages, etc). As a result, hardly any two instrument configurations in different laboratories correspond completely with each other. This makes handling very demanding for operators, maintenance personnel, and service engineers.
Methods implemented on these heterogeneous platforms (e.g. instruments from various vendors) are in turn characterized by a very considerable number of variables, e.g. chromatographic gradients, labelling patterns of internal standards, purity of solvents, dead volume of flow paths, etc.
Taken together, the variety of assays referring to an identical analyte (such as tacrolimus or testosterone) is enormous, with an almost astronomical combinatorial complexity.
However, method publications are still traditionally written more or less in a case report approach: the feasibility and performance of a method realization is demonstrated for one individual system configuration. It is usually not clear which features are really essential for the method and which features can be variable between different implementations – and which second implementation can still be considered ‘the same’ method. This means that the question of the true ‘identity’ of a method has not yet been deepened by application notes or publications in scientific journals; thus the level of abstraction required here is missing.
In an attempt to standardize the description of MS/MS-based methods, we selected a set of 35 characteristics that are defined as essential for a method (see Table 1) [7], for example, main approach of sample preparation (e.g. protein precipitation with acetonitrile), main technique of ionization (e.g. electrospray ionization in negative mode); molecular structure of the internal standard; mass transitions; calibration range. In addition, we define 15 characteristics of a method that cannot or should not be realistically standardized in time and space (examples: manufacturer and brand of the MS detector; dead volume of the flow path; lot of analytical columns and solvents). These characteristics – identified as variable – should be documented in the internal report files.
We found it feasible to describe several exemplary MS/MS methods using this scheme and a corresponding matrix. On the basis of this matrix, the method transfer to different platforms and laboratories will be much easier and more reliable. Specifying the identity of a method in the proposed way has the essential advantage that a method revalidation can be transparently triggered by defined criteria, e.g. the use of a novel internal standard with a different labelling pattern.
The proposed scheme for method description may also be the basis of a comprehensive traceability report for any result obtained by an MS-based method in the clinical laboratory. Standardization of batch release (Table 2) While today’s routine analyser platforms essentially provide unambiguous final results for each sample, the process of generating quantitative results from primary data in MS is open and transparent. Primary data in MS are the peak areas of the target analyte observed in diagnostic samples. In addition to these primary data, a range of metadata is provided (e.g. internal standard area, peak height-to-area, peak skewness, qualifier peak area; metadata related to analytical batches, e.g. coefficient of variation (CV) of internal standard areas). This transparency and abundance of data is a cornerstone of the high potential reliability of MS-based assays and therefore their interpretation is very important [8, 9].
However, the evaluation of this metadata – related to individual samples and batches – is nowadays done very heterogeneously from laboratory to laboratory [10]; this applies to LDTs as well as to commercially available kit products. The structure of analytical batches is also very variable and there is no generally accepted standard (number and sequence of analysis of calibration samples in relation to patient and quality control samples, blank injections, zero samples, etc).
While the validation of methods – which is performed before a method is introduced into the diagnostic routine – is discussed in detail in the literature (and in practice), the procedures applied to primary data before release for laboratory reporting have not yet been standardized. Validation is generally defined as the process of testing whether predefined performance specifications are met. Therefore, quality control and release of analytical batches and patient results should also be considered a process of validation, and criteria for the acceptance or rejection of results should be predefined.
A three-step approach to validation, covering the entire life cycle of methods in the clinical laboratory, can be conceptualized: dynamic validation should integrate validation of methods, validation of analytical batches and validation of individual test readings. We believe that standardization of this process of batch and sample result validation and release is needed as a guide for developers of methods, medical directors, and technicians.
In a recent article published in Clinical Mass Spectrometry [11], we propose a list of characteristics that should be considered for batch and sample release. In this article we only mention figures for merits and issues to be addressed and do not claim to have specific numerical acceptance criteria. Therefore, this generic list of items is intended as a framework for the development of an individual series and batch validation plan in a laboratory. Furthermore, we consider this list to be a living document, subject to further development and standardization as the field matures.
We believe that it is essential to include basic batch and sample release requirements as essential characteristics in the description of a method [7]. Therefore, we believe that efforts to standardize method description and batch/sample release should be synergistically linked to facilitate the use of MS in routine laboratories.
The approach proposed to clinical MS in these two companion articles [7, 11] can be the basis for discussion and eventually for the development of official standards for these areas by the Clinical and Laboratory Standards Institute (CLSI) and/or International Organization for Standardization (ISO). We believe that these documents can provide a solid basis for internal and external audits of LC-MS/MS-based Quality Control April/May 2020 9 | LDTs, which will become particularly relevant in the context of the IVD Regulation 746 in the European Union [12].
Both approaches – standardized description of MS methods and standardization of batch release – aim at implementing methodological traceability. This corresponds to the analytical standardization and metrological traceability of measurements to higher order reference materials [13, 14]. Future perspectives In the future, a commercialization of MS-based black-box instruments on a larger scale is expected. However, LC-MS/MS will remain a critical technique for LDTs, and the flexibility of MS to develop tests on demand – independent of the IVD industry on fully open LC-MS/MS platforms – will remain a key pillar of laboratory medicine.
Both publications, which this article puts into context [7, 11], have been published in Clinical Mass Spectrometry, the first and only international journal dedicated to the application of MS methods in diagnostic tests including publications on best practice documents. Both articles are freely available.
Clinical Mass Spectrometry is the official journal of MSACL (The Association for Mass Spectrometry: Applications to the Clinical Laboratory; www.msacl.org). MSACL organizes state-of-the-art congresses that focus on translating MS from clinical research to diagnostic tests (i.e. bench to clinic).
In summary, we advocate innovative approaches to methodological standardization of LC-MS/MS methods to master the complexity of this powerful technology and to facilitate and promote its safe application in clinical laboratories worldwide. The authors Michael Vogeser*1 MD, Judy Stone2 PhD, Alan Rockwood3 PhD 1 Hospital of the University of Munich (LMU), Institute of Laboratory Medicine, Munich, Germany University of California, San Francisco Medical Center, Laboratory Medicine, Parnassus Chemistry, San Francisco, CA, USA 3 Rockwood Scientific Consulting, Salt Lake City, UT, USA
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The UK-based Covid-19 Volunteer Testing Network launched April 9 to provide essential additional testing capacity to front-line workers. The project, started by Mike Fischer CBE, helps small laboratories convert to run critical antigen testing and identify Covid-19 cases among local healthcare workers – at no cost to Government.
The UK has thousands of small laboratories with the right equipment, personnel and processes to run Covid-19 testing. Although some of the critical RT-PCR machines in university and healthcare settings have already been requisitioned by central Government, thousands of others are currently sitting idle in small, ‘long-tail’ facilities up and down the United Kingdom.
Fischer set up SBL, a non-profit medical research laboratory in Oxfordshire, which is already running 250-500 tests a week for 10 GP surgeries in the local area.
“Although our facility is small – with just three full-time staff, two containment hoods and two real-time machines – we were quickly able to convert to Covid-19 testing using the Centre for Disease Control protocols and are now running up to 500 tests a week for the staff at 10 local GP surgeries on a same-day basis,” said Fischer.
“If other labs could join the effort we could quickly scale to providing tens of thousands of tests a day in complement to the central program.”
“If we are going to beat this pandemic, we need to employ every resource we can to make sure that our essential health care workers can go to work safely. Even at our small facility, we have been able to run up to 500 tests a week for NHS staff on a same-day basis. By creating an emergency network of volunteer laboratories like ours across the UK, we can quickly and efficiently create the capacity we need to deliver tens of thousands of additional tests every day.”
The Covid-19 Volunteer Testing Network is being coordinated on an entirely voluntary basis and is looking for further labs to join the effort. “We hope existing equipment can be used in situ with qualified staff volunteering to conduct the tests. We are able to provide guidance, protocols, documentation and reporting,” Fischer added.
The Fischer Family Trust has also made £1 million in funding available to support the purchase of consumables for the tests if labs are unable to cover these.
For more information about the Covid-19 Volunteer Testing Network, visit: www.covid19-testing.org
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NanoPass is sharing its proprietary MicronJet microneedle device with leading vaccine and immunotherapy companies around the world to assist in development of a Covid-19 vaccine.
The NanoPass device targets immune cells of the skin by harnessing the skin’s potent immune system to improve vaccines and/or to dramatically reduce the dose while achieving the same immunity.
“The human skin is our first layer of defence against many infectious diseases,” says Yotam Levin, MD, CEO of NanoPass. “The skin contains specialized Dendritic Cells that process and induce strong immune responses – that’s why microneedle injections enable reduction of vaccine doses by five-fold, thereby reducing overall cost, required capacity and production time. We believe a reliable injection into the skin is critical for successful activation of broad and effective immune responses, which should be explored for most injectable vaccines.”
The company’s technology is supported by more than 55 completed/ongoing clinical studies with various vaccines and vaccine platforms, including H1N1, H5N1 and live attenuated VZV vaccine, that have shown improved immunogenicity and significant dose-sparing. Pre-clinical evidence with mRNA and DNA vaccines showed promising results.
NanoPass has previously supported US CDC in a Phase 3 infant polio vaccination trial; with ITRC on PPD skin testing; in Type 1 Diabetes immunotherapy; and supported NIAID with devices to evaluate immunogenicity of a pandemic flu vaccine; and multiple vaccine pharma.
NanoPass Technologies flagship product, the 0.6 mm MicronJet, is the first true (<1 mm) microneedle to receive FDA clearance as an intradermal delivery device for substances approved for delivery below the surface of the skin. It is supported by extensive clinical data and regulatory approvals in most major markets including the US, Europe, China and Korea.
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Fujirebio Europe has received the CE mark for the molecular IVD assay iAMP Covid-19 Detection Kit from its partner Atila Biosystems. The qualitative detection kit is based on real-time fluorescent reverse transcription isothermal amplification, eliminating the need for RNA extraction.
The detection kit was also granted Emergency Use Authorization by the US Food and Drug Administration on April 10.
The iAMP COVID-19 Detection Kit can be run on a Real-Time PCR PowerGene 9600 Plus or any other qPCR automate capable of measuring fluorescence in FAM/HEX channel in real-time.
The new iAMP COVID-19 molecular assay complements the existing panel of biomarkers available on the LUMIPULSE® G System for infection (PCT, Ferritin), inflammation (IL-6) and epithelial lung injury (KL-6) to predict disease severity in patients infected with SARS-CoV-2.
Products from Atila Biosystems are available through Fujirebio’s European affiliates and through a large portion of Fujirebio’s existing or new European distribution network.
For more information, visit: www.fujirebio.com/en/contact
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BIOHIT Healthcare is distributing test kits for the diagnosis of both current and past COVID-19 infections to help in the fight against coronavirus in the UK. The new product line includes the MutaPLEX® Coronavirus kit from Immundiagnostik AG (IDK) – a real-time RT-PCR assay to screen for infected individuals – and Epitope Diagnostics Inc’s (EDI’s) immunodiagnostic tests for IgM and IgG COVID-19 antibodies, to detect past infections.
The IDK MutaPLEX coronavirus screening assay allows the detection of SARS-CoV-2 viral RNA in a variety of biological specimens, especially nasal/throat swabs. This real time RT-PCR kit contains all the reagents, primers and dual-labelled probes required for the amplification and simultaneous differentiation of RNA from SARS-CoV-2 and other betacoronaviruses, as well as house-keeping genes designed to prevent false negative results due to insufficient sample collection or transport problems.
EDI’s Novel Coronavirus COVID-19 ELISA kits provide qualitative detection of antibodies in patient serum, indicating a past COVID-19 infection. The IgM assay provides the earliest immunodiagnostic indication of an infection, while the IgG test can be used to aid detection and provide an indication of long-term immunological response, making it particularly useful in cases where clustering is suspected or differential diagnosis is required.
These tests extend and complement BIOHIT’s repertoire of diagnostic kits for gastroenterology, aiding the evaluation of patients with both GI and upper respiratory complaints, as COVID-19 may include stomach and bowel symptoms in some cases. Inflammatory bowel disease patients being treated with immunosuppressive agents should also be considered at high risk for COVID-19, making differential diagnosis essential.
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UK-based Avacta Group, the developer of Affimer biotherapeutics and reagents, will collaborate with US-based Adeptrix to develop a high throughput Covid -19 antigen test using Adeptrix’s proprietary bead-assisted mass spectrometry (BAMS) platform.
The Affimer-based BAMS coronavirus antigen test that will provide clinicians with a significant expansion of the available testing capacity for Covid-19 infection in hospitals.
Adeptrix’s novel BAMS platform combines enrichment of the sample to improve sensitivity with the power of mass-spectrometry to improve specificity. Hundreds of samples per day can be analysed by a single technician using BAMS, exceeding the capacity of single PCR machine, making BAMS a very attractive high throughput technique for Covid-19 screening in the clinical setting.
The diagnostic test will allow hospitals around the world to utilise their existing installed base of mass spectrometers that are not currently used for Covid-19 testing, thus contributing significantly to the increase in global testing capacity. Avacta’s recently developed Affimer reagents that bind the SARS-COV-2 spike protein will be used to provide the capture and enrichment of the virus particle from the sample which could be saliva, nasopharyngeal swabs or serum.
The companies are aiming to have a BAMS test ready for clinical validation, regulatory approval and manufacturing in June. Adeptrix and Avacta are already in discussion with large-scale manufacturing partners to rapidly deploy this new high throughput test.
Dr Alastair Smith, Chief Executive Officer of Avacta Group, commented: “We believe that the BAMS test will be hugely attractive as an adjunct to PCR testing because it uses laboratory equipment that is already in hospital labs but not currently used for Covid-19 testing so it provides incremental testing capacity.
I have made it clear that we intend to partner the SARS-COV-2 spike protein Affimer reagents with several select companies to support antigen test development on multiple diagnostic test platforms. This will contribute most effectively to the urgent need to increase antigen testing capacity globally and maximise the commercial return to Avacta. Adeptrix is one example of this and other discussions are underway. I look forward very much to further updating the market in the near future.”
Dr. Jeffrey C. Silva, Director of Product Development, Adeptrix Corporation commented: “Mass spectrometry can enhance the diagnostic utility of immunoassays, as it is capable of monitoring both existing and emerging viral strains by accurately measuring the molecular components of the virus. BAMS provides an ideal multiplexing platform to obtain higher specificity for monitoring Covid-19 infection.”
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UK-based iPSC (induced pluripotent stem cells) disease modelling company DefiniGEN has identified iPSC-derived intestinal organoids that could be used to help structure in vitro studies of the biology of SARS-CoV-2 infection across cohorts of multiple patients.
While SARS-CoV-2 primarily targets the respiratory system, studies have shown that it also infects and multiplies within the intestinal epithelium. IPSC-derived organoids exhibit characteristics that closely mimic the in vivo intestinal epithelium, making them a valuable surrogate model for studying the virus.
The company says their iPSC-derived intestinal organoids provide a unique in vitro system to model the human intestine. The organoids display a polarized epithelium and harbour a mixture of cell types normally present in the primary intestinal epithelium barrier in vivo, including goblet cells, Paneth cells, enterocytes, LRG5+ stem cells, and enteroendocrine cells. The organoids polarise, form crypt structures and grow villi at the apical surface, and are shown to secrete mucus in a similar manner to primary human gut tissue.
DefiniGEN points out that several studies have proven that angio-tensin-converting enzyme 2 (ACE2) expression in host cells is required for SARS-CoV-2 recognition and infection. Activity of membrane proteases such as TMPRSS2 cleaves the coronavirus’ Spike protein and facilitates the membrane fusion with the host cell. Human intestine is one of the few human tissues with high expression of both ACE2 and TMPRSS2 therefore is a good candidate to study Covid-19 and the mechanisms of the SARS-CoV-2 infection.
Additionally, DefiniGEN have a platform to generate various patient-derived intestinal models which could support population studies, using many different donors with diverse ethnic profiles.
Such studies are useful as there is growing evidence that ethnic differences are a major factor in patients showing a severe response to Covid-19.
DefiniGEN’s differentiation platform is optimized to enable successful generation of intestinal organoids from a diverse range of patients. Patient skin fibroblasts or PBMCs can first be reprogrammed to iPSC, and then differentiated to produce mature intestinal organoids which carry the original patient genetics, and so manifest a gut model specific to that donor.
For more information, visit www.definigen.com/products/intestinal/covid-19
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Siemens Healthineers announced late May that it is now shipping worldwide its laboratory-based total antibody test to detect the presence of SARS-CoV-2 IgM and IgG antibodies in blood. The test received the CE mark and data has demonstrated 100 percent sensitivity and 99.8 percent specificity. The total antibody test allows for identification of patients who have developed an adaptive immune response, which indicates recent infection or prior exposure.
The US FDA has issued an Emergency Use Authorization (EUA) for its laboratory-based total antibody test.
Siemens says it is prepared to ramp up production as the pandemic evolves with capacity exceeding 50 million tests per month across its platforms starting in June.
The antibody test is now available on the largest installed base in the U.S. and one of the largest in the world with 20,000 Siemens Healthineers systems installed worldwide. This includes the Atellica Solution immunoassay analyser, which can run up to 440 tests per hour and enables a result in just 10 minutes. By detecting both IgM and IgG antibodies, the test provides a clearer clinical picture over a longer period of time as the disease progresses.
The antibody test also is available on the company’s installed base of ADVIA Centaur XP and XPT analysers, which can test up to 240 samples per hour, with a result in 18 minutes.
Importantly, the test detects antibodies to a key spike protein on the surface of the SARS-CoV-2 virus, which binds the virus to cells with a distinct human receptor found in lungs, heart, multiple organs and blood vessels. Studies indicate that certain (neutralizing) antibodies to the spike protein can disarm SARS-CoV-2, presumably by interfering with the ability of the virus to bind, penetrate and infect human cells. Multiple potential vaccines in development for SARS-CoV-2 include the spike protein within their focus.
https://clinlabint.com/wp-content/uploads/sites/2/2020/08/CLI-siemens_antibody-scaled.jpg141625603wmediahttps://clinlabint.com/wp-content/uploads/sites/2/2020/06/clinlab-logo.png3wmedia2020-08-26 09:31:372021-01-08 11:07:50Siemens starts worldwide shipping of total antibody test for COVID-19
Avacta Group plc, the developer of Affimer biotherapeutics and reagents, has started shipping Affimer reagents for COVID-19 antigen testing to its diagnostic test development partners.
The Group recently reported that it had generated multiple, highly specific Affimer reagents that bind the SARS-COV-2 viral antigen and do not cross-react with SARS, MERS and other closely related coronaviruses. These Affimer reagents will be used to develop a point-of-care saliva based COVID-19 antigen test strip by Cytiva (formerly GE Healthcare Life Sciences) for CE marking in Europe and FDA approval in the United States.
The Affimer reagents have been manufactured by Avacta in the quantities required for test development and are being sent to Cytiva. The reagents are also being provided to Adeptrix with whom Avacta has announced that it will develop a COVID-19 laboratory test to run on hospital mass spectrometers using Adeptrix’s proprietary BAMS assay platform.
The Affimer reagents have been studied further by Avacta and this has shown that there are Affimer reagents that can work in pairs, both binding to the spike protein at the same time. This allows tests to be developed that detect both the intact virus particle and the detached spike proteins which become separated from the virus particle during the development of the COVID-19 disease, which may also be important in monitoring disease progression.
Cytiva and Avacta will now work to develop rapid test strips for the detached spike protein and for the intact virus particle. Adeptrix is working to develop a prototype BAMS test. Both of these tests will indicate whether a person has the infection at that moment.
https://clinlabint.com/wp-content/uploads/sites/2/2020/06/clinlab-logo.png003wmediahttps://clinlabint.com/wp-content/uploads/sites/2/2020/06/clinlab-logo.png3wmedia2020-08-26 09:31:372021-01-08 11:07:50Avacta ships SARS-COV-2 Affimer reagents to Cytiva and Adeptrix for diagnostic test development
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Consider moving average quality control when internal control is insufficient or inefficient – the time is now!
, /in E-News /by 3wmediaby Dr Huub H. van Rossum
Recently, significant improvements have been made in understanding and applying moving average quality control (MA QC) that enable its practical implementation. These include the description of new and laboratory-specific MA QC optimization and validation methods, the online availability thereof, insights into operational requirements, and demonstration of practical implementation.
Introduction
Moving average quality control (MA QC) is the process of algorithmically averaging obtained test results and using that average for (analytical) quality control purposes. MA QC is generally referred to as patient-based real-time quality control (PBRTQC) because it is one of various methods (e.g. limit checks, delta checks, etc) that use patient results for (real-time) quality control. MA QC was first described over half a century ago as ‘average of normals’ [1]. Since then, it has evolved into a more general MA QC concept not necessarily based on using mean calculations of the obtained ‘normal’ test results [2]. Although MA QC has been available for a few decades, its adoption by laboratories has been limited due to the complexity of setting up the necessary procedures, operational challenges and a lack of evidence to justify its application and demonstrate its value. During the past 5|years, however, significant improvements have been made in the field of MA QC, and research studies have addressed all these issues. Consequently, true practical application of validated MA QC procedures to support analytical quality control in medical laboratories is now possible. Furthermore, the recent improvements may well change the way we perform daily analytical quality control in medical laboratories in the near future.
MA QC optimization and validation
The recent significant improvements in the field of MA QC include, first and foremost, the description of new methods to design and optimize laboratory-specific MA QC procedures and to enable validation of their actual error-detection performance [2–5]. These methods use realistic MA QC simulations based on laboratory-specific datasets and thus provide objective insights into MA QC error detection [2]. To enable practical implementation, the requirement that the number of MA QC alarms must be manageable is now acknowledged as essential and has been fulfilled when setting up MA QC [2, 6]. The newly developed methods use a novel metric to determine the error-detection performance: that is, the mean or median number of test results needed for error detection. One of the new methods presents these simulation results in bias-detection curves so that the optimal MA QC procedure can be selected, based on its overall error-detection performance [5]. An example of a bias-detection curve and its application is presented in Figure 1. After selecting the optimal MA QC settings, an MA validation chart can be used to obtain objective insights into the overall error-detection performance and the uncertainty thereof. Therefore, this chart can be seen as a validation of the MA QC procedure. An example of an MA validation chart is presented in Figure 2 and shows that the MA QC procedure will almost always (with 97.5% probability) detect a systematic error of −4% (or larger negative errors) within 20 test results.
Importantly, this method has become available to laboratories via the online MA Generator application, enabling them to design their own optimized and validated MA QC procedures [7]. Laboratories can now upload their own datasets of historical results, study potential MA QC settings using this simulation analysis and obtain their own laboratory-specific MA QC settings and MA validation charts. Several laboratories have demonstrated that this tool has enabled them to obtain relevant MA QC settings and thus implement MA QC [8, 9].
Integration of MA QC with internal QC
Measurement of internal quality control (iQC) samples is still the cornerstone of analytical quality control as performed in medical laboratories. For many tests, iQC alone is sufficient to assure and control the quality of obtained test results. For some tests, however, iQC itself is insufficient. The reasons for this are related to certain fundamental characteristics of iQC that include: lack of available (stable) control materials, its scheduled character, the risk of using non-commutable control samples and tests with a sigma metric score of ≤4. For several reasons, PBRTQC or, more specifically, MA QC is a particularly valuable and powerful way to support quality assurance in such cases.
First, if no (stable) QC materials are available it is impossible, or it becomes complicated, to use iQC. This is, for example, relevant for the erythrocyte sedimentation rate, serum indices or hemocytometry tests including erythrocyte mean corpuscular volume in particular. MA QC is possible as long as patient results are available. Second, the scheduled character of iQC becomes a limitation and a risk when temporary assay failures or rapid onset of critical errors occur between scheduled iQC. Because a new MA QC value can be calculated for each newly obtained test result, MA QC can be designed as a continuous and real-time QC tool. In this context, detection of temporary assay failure by MA QC between scheduled iQC has been demonstrated for a sodium case [10], and several examples of MA QC detection of rapid onset of critical errors have been published for both chemistry and hematological tests [11]. Third, because PBRTQC methods such as MA QC use obtained patient results, by design there is no commutability issue. Fourth, and finally, for some tests iQC is intrinsically limited in its ability to detect relevant clinical errors, due to the low ratio of biological variations to analytical variations, as reflected in low sigma metric values. Such tests require frequent iQC analysis and application of stringent control rules. However, even with such an intensive and strict iQC set-up, the probability of detecting clinically relevant errors remains limited [12]. In contrast, MA QC has the best error-detection performance for tests with a low sigma value [13].
For all these reasons, MA QC is ideal for supplementing analytical quality control by iQC. Recently, an approach was presented that integrated MA QC into the QC plan when iQC was found to be insufficient [9]. This approach was based on first determining whether one of the abovementioned iQC limitations applied to a test. If so, then iQC alone was considered insufficient and MA QC was studied, using the online MA Generator tool (www.huvaros.com) to obtain optimal MA QC settings and MA QC procedures to support the analytical quality control [7, 9]. The MA QC error-detection performance was validated using MA validation charts. These latter insights into MA QC error detection also enabled iQC measurements to be reduced. The MA QC procedures alone provided significant error-detection performance, so running iQC measures multiple times a day would add only limited error-detection performance. Therefore, it was decided to run the iQC only once a day and add the obtained MA QC procedures to the QC plan.
Others have taken this a step further and studied MA QC not only for tests with limited iQC performance but also for a much larger test selection, in order to reduce the number of iQC measures and more efficiently schedule and apply iQC [4]. This approach has been shown to be successful for a large commercial laboratory with high production numbers. Since the MA QC error-detection performance improves with an increasing number of test results and benefits from a small number of pathological test results, this approach may be particularly valuable to the larger commercial laboratories. For such an approach, the key is objective insights into the error-detection performance of MA QC procedures such as obtained using MA validation charts.
Implementation and application of MA QC for real-time QC in medical laboratories
The final aspect in which there have been significant improvements in recent years relates to the practical application of MA QC in medical laboratories. Recently, an International Federation of Clinical Chemistry and Laboratory Medicine working group was founded that summarized medical laboratories’ experiences of practically applying MA QC and formulated several recommendations for both MA QC software suppliers and medical laboratories that are working on, or are interested in, implementation of MA QC [14, 15]. Also, a step-by-step roadmap has recently been published to enable MA QC implementation [9]. The first two steps of this roadmap – i.e. selection of tests and obtaining MA QC settings for them – were discussed in the previous two paragraphs.
The next step would be to set up and configure the software used to implement MA QC in medical laboratories. If you are interested in applying MA QC in your laboratory, it is important to review the available software (e.g. analyser, middleware, LIS, third party) and to decide which will be used to run and apply MA QC. Your decision depends not only on the availability of suitable software in or for the laboratory, but also on the actual MA QC functionality present in the software packages.
The minimum software features that are necessary to enable practical implementation have been formulated [2, 15]. In my view, key elements would be that the software supports: exclusion of specified samples (non-patient materials, QC results, extreme results, etc), calculation of relevant MA QC algorithms, applying SD-based as well as non-statistical control limits (plain lower and upper control limits), proper real-time alarming and – depending on the MA QC optimization method – presentation of MA QC in a Levey–Jennings or accuracy graph. Figure 3 presents an example of MA QC in an accuracy graph as operated for real-time QC in my laboratory. To enable effective implementation of MA QC, all of these software features should be configured.
The final implementation step I wish to address here is the design of laboratory protocols for working up MA QC alarms, which determines the extent to which an error detected by an MA QC alarm is acknowledged. An important requirement is that all MA QC alarms should be worked up by means of this protocol.
As previously indicated, because MA QC can generate many more QC results and alarms than iQC, a critical requirement of every MA QC procedure is a manageable number of alarms. As a result, when an MA QC alarm occurs there is a reasonable chance of detecting error.
A first common action as part of the MA QC alarm protocol would be to run iQC. This provides a quick insight into the size of the error and enables rapid confirmation of large errors. As a second step, re-running of recently analysed samples (in addition to running iQC) enables temporary assay failures to be detected and can confirm or exclude errors not necessarily detectable by iQC. Also, finally, a review of recently analysed test results to identify a pre-analytical cause or a single patient with extreme but valid test results is often very useful as part of the MA QC alarm protocol. All these aspects have recently been discussed in greater detail [10, 14].
Conclusions
Altogether, the recent developments in the field of PBRTQC and, more specifically, MA QC now – finally – enable true practical implementation of MA QC in medical laboratories and allow more effective and efficient QC plans to be designed.
The authors
Huub H. van Rossum1,2 PhD
1 Department of Laboratory Medicine, The Netherlands Cancer Institute, Amsterdam, The Netherlands
2 Huvaros, Amsterdam, The Netherlands
E-mail: h.v.rossum@nki.nl
Better health care through mass spectrometry – better mass spectrometry through standardization
, /in E-News, Editors' Picks /by 3wmediaby Prof. Michael Vogeser, Dr Judy Stone and Prof. Alan Rockwood
While analytical standardization and metrological traceability are well-defined terms, ‘methodological standardization’ in clinical mass spectrometry is still in a developing stage. We propose a framework that facilitates the widespread implementation of this highly complex and very powerful technology and is based on two pillars – standardization of the description of LC-MS/MS methods and standardization of the release of clinical test results as a three-step sequence of method validation, batch validation and validation of individual measurements.
Mass spectrometry in the clinical laboratory
Mass spectrometry (MS)-based methods now play an important role in many clinical laboratories worldwide. To date, areas of application have focused especially on screening for hereditary metabolic diseases, therapeutic drug monitoring, clinical toxicology and endocrinology. In fact, these techniques offer significant advantages over immunoassays and photometry as basic standard technologies in clinical chemistry: high analytical selectivity through true molecular detection; wide range of applications without the need for specific molecular features (as in UV detection or specific epitopes); high multiplexing capacity and information-rich detection; and, in many cases, matrixindependent analyses, thanks to the principle of isotope dilution [1].
Various MS technologies – in particular tandem MS (MS/MS-coupling with molecular fragmentation), time-of-flight (TOF) MS and Orbitrap-MS – with front-end fractionation technologies such as HPLC or UPLC potentially allow very reliable analysis, but the technology itself is no guarantee of this: these techniques have a very high complexity and a wide range of potential sources of error [2] which require comprehensive quality assurance [3–5]. Indeed, the high degree of complexity is still the main hurdle for the application of MS in the special environment of clinical laboratories. Specific challenges of this type of laboratory – in contrast to research and development laboratories – include: heterogeneous mix of staff qualifications; requirement for maximum handling safety when operating a large number of analysis platforms; work around the clock; and direct impact on the outcome of the individual patient.
Indeed, after more than two decades of commercial availability of LC-MS/MS instruments, their application in a global perspective has remained very limited. The translation of MS into fully automated ‘black box’ instruments is underway, but still far from being realized on a large scale [6], with laboratory developed tests (LDTs) still dominating the field of clinical MS applications. Kit solutions for specific analytes provided by the in vitro diagnostics (IVD) industry are becoming increasingly available, but their application also requires a very high level of skills and competence from laboratories.
Two main differences of MS-based LDTs as opposed to standard ‘plug-and-play’ analysis systems in today’s clinical laboratories can be identified: first, the high heterogeneity of device configurations and second, the handling of large amounts of data, from sample list structures to technical metadata analysis.
In fact, the random access working mode is now so widespread in all clinical laboratories that the ‘analytical batch’ is no longer standard in laboratories. In the same way, modern analytical instruments no longer challenge the end users with extensive metadata (such as reaction kinetics or calibration diagrams). To achieve the goal of making the extraordinary and disruptive analytical power of MS fully usable for medicine to an appropriate extent, approaches to master the heterogeneity of platform configurations and to regulate the handling of batches and metadata are urgently needed – and standardization efforts seem to be crucial in this context.
Standardization of the method description
IVD companies manufacture many different instrument platforms, but each of these platforms is very homogeneous worldwide and is produced in large quantities for years. In contrast, MS platforms in clinical laboratories have to be individually assembled from a very large number of components from many manufacturers (sample preparation modules, autosamplers, high performance pumps, switching valves, chromatography columns, ion sources, mass analysers, vacuum systems, software packages, etc). As a result, hardly any two instrument configurations in different laboratories correspond completely with each other. This makes handling very demanding for operators, maintenance personnel, and service engineers.
Methods implemented on these heterogeneous platforms (e.g. instruments from various vendors) are in turn characterized by a very considerable number of variables, e.g. chromatographic gradients, labelling patterns of internal standards, purity of solvents, dead volume of flow paths, etc.
Taken together, the variety of assays referring to an identical analyte (such as tacrolimus or testosterone) is enormous, with an almost astronomical combinatorial complexity.
However, method publications are still traditionally written more or less in a case report approach: the feasibility and performance of a method realization is demonstrated for one individual system configuration. It is usually not clear which features are really essential for the method and which features can be variable between different implementations – and which second implementation can still be considered ‘the same’ method. This means that the question of the true ‘identity’ of a method has not yet been deepened by application notes or publications in scientific journals; thus the level of abstraction required here is missing.
In an attempt to standardize the description of MS/MS-based methods, we selected a set of 35 characteristics that are defined as essential for a method (see Table 1) [7], for example, main approach of sample preparation (e.g. protein precipitation with acetonitrile), main technique of ionization (e.g. electrospray ionization in negative mode); molecular structure of the internal standard; mass transitions; calibration range. In addition, we define 15 characteristics of a method that cannot or should not be realistically standardized in time and space (examples: manufacturer and brand of the MS detector; dead volume of the flow path; lot of analytical columns and solvents). These characteristics – identified as variable – should be documented in the internal report files.
We found it feasible to describe several exemplary MS/MS methods using this scheme and a corresponding matrix. On the basis of this matrix, the method transfer to different platforms and laboratories will be much easier and more reliable. Specifying the identity of a method in the proposed way has the essential advantage that a method revalidation can be transparently triggered by defined criteria, e.g. the use of a novel internal standard with a different labelling pattern.
The proposed scheme for method description may also be the basis of a comprehensive traceability report for any result obtained by an MS-based method in the clinical laboratory.
Standardization of batch release (Table 2)
While today’s routine analyser platforms essentially provide unambiguous final results for each sample, the process of generating quantitative results from primary data in MS is open and transparent. Primary data in MS are the peak areas of the target analyte observed in diagnostic samples. In addition to these primary data, a range of metadata is provided (e.g. internal standard area, peak height-to-area, peak skewness, qualifier peak area; metadata related to analytical batches, e.g. coefficient of variation (CV) of internal standard areas). This transparency and abundance of data is a cornerstone of the high potential reliability of MS-based assays and therefore their interpretation is very important [8, 9].
However, the evaluation of this metadata – related to individual samples and batches – is nowadays done very heterogeneously from laboratory to laboratory [10]; this applies to LDTs as well as to commercially available kit products. The structure of analytical batches is also very variable and there is no generally accepted standard (number and sequence of analysis of calibration samples in relation to patient and quality control samples, blank injections, zero samples, etc).
While the validation of methods – which is performed before a method is introduced into the diagnostic routine – is discussed in detail in the literature (and in practice), the procedures applied to primary data before release for laboratory reporting have not yet been standardized. Validation is generally defined as the process of testing whether predefined performance specifications are met. Therefore, quality control and release of analytical batches and patient results should also be considered a process of validation, and criteria for the acceptance or rejection of results should be predefined.
A three-step approach to validation, covering the entire life cycle of methods in the clinical laboratory, can be conceptualized: dynamic validation should integrate validation of methods, validation of analytical batches and validation of individual test readings. We believe that standardization of this process of batch and sample result validation and release is needed as a guide for developers of methods, medical directors, and technicians.
In a recent article published in Clinical Mass Spectrometry [11], we propose a list of characteristics that should be considered for batch and sample release. In this article we only mention figures for merits and issues to be addressed and do not claim to have specific numerical acceptance criteria. Therefore, this generic list of items is intended as a framework for the development of an individual series and batch validation plan in a laboratory. Furthermore, we consider this list to be a living document, subject to further development and standardization as the field matures.
We believe that it is essential to include basic batch and sample release requirements as essential characteristics in the description of a method [7]. Therefore, we believe that efforts to standardize method description and batch/sample release should be synergistically linked to facilitate the use of MS in routine laboratories.
The approach proposed to clinical MS in these two companion articles [7, 11] can be the basis for discussion and eventually for the development of official standards for these areas by the Clinical and Laboratory Standards Institute (CLSI) and/or International Organization for Standardization (ISO). We believe that these documents can provide a solid basis for internal and external audits of LC-MS/MS-based Quality Control April/May 2020 9 | LDTs, which will become particularly relevant in the context of the IVD Regulation 746 in the European Union [12].
Both approaches – standardized description of MS methods and standardization of batch release – aim at implementing methodological traceability. This corresponds to the analytical standardization and metrological traceability of measurements to higher order reference materials [13, 14].
Future perspectives
In the future, a commercialization of MS-based black-box instruments on a larger scale is expected. However, LC-MS/MS will remain a critical technique for LDTs, and the flexibility of MS to develop tests on demand – independent of the IVD industry on fully open LC-MS/MS platforms – will remain a key pillar of laboratory medicine.
Both publications, which this article puts into context [7, 11], have been published in Clinical Mass Spectrometry, the first and only international journal dedicated to the application of MS methods in diagnostic tests including publications on best practice documents. Both articles are freely available.
Clinical Mass Spectrometry is the official journal of MSACL (The Association for Mass Spectrometry: Applications to the Clinical Laboratory; www.msacl.org). MSACL organizes state-of-the-art congresses that focus on translating MS from clinical research to diagnostic tests (i.e. bench to clinic).
In summary, we advocate innovative approaches to methodological standardization of LC-MS/MS methods to master the complexity of this powerful technology and to facilitate and promote its safe application in clinical laboratories worldwide.
The authors
Michael Vogeser*1 MD, Judy Stone2 PhD, Alan Rockwood3 PhD
1 Hospital of the University of Munich (LMU), Institute of Laboratory Medicine, Munich, Germany
University of California, San Francisco Medical Center, Laboratory Medicine, Parnassus Chemistry, San Francisco, CA, USA
3 Rockwood Scientific Consulting, Salt Lake City, UT, USA
* Corresponding author
E-mail: Michael.Vogeser@med.uni-muenchen.de
Volunteer laboratory network launched in UK to expand Covid-19 testing
, /in Corona News, E-News /by 3wmediaThe UK-based Covid-19 Volunteer Testing Network launched April 9 to provide essential additional testing capacity to front-line workers. The project, started by Mike Fischer CBE, helps small laboratories convert to run critical antigen testing and identify Covid-19 cases among local healthcare workers – at no cost to Government.
The UK has thousands of small laboratories with the right equipment, personnel and processes to run Covid-19 testing. Although some of the critical RT-PCR machines in university and healthcare settings have already been requisitioned by central Government, thousands of others are currently sitting idle in small, ‘long-tail’ facilities up and down the United Kingdom.
Fischer set up SBL, a non-profit medical research laboratory in Oxfordshire, which is already running 250-500 tests a week for 10 GP surgeries in the local area.
“Although our facility is small – with just three full-time staff, two containment hoods and two real-time machines – we were quickly able to convert to Covid-19 testing using the Centre for Disease Control protocols and are now running up to 500 tests a week for the staff at 10 local GP surgeries on a same-day basis,” said Fischer.
“If other labs could join the effort we could quickly scale to providing tens of thousands of tests a day in complement to the central program.”
“If we are going to beat this pandemic, we need to employ every resource we can to make sure that our essential health care workers can go to work safely. Even at our small facility, we have been able to run up to 500 tests a week for NHS staff on a same-day basis. By creating an emergency network of volunteer laboratories like ours across the UK, we can quickly and efficiently create the capacity we need to deliver tens of thousands of additional tests every day.”
The Covid-19 Volunteer Testing Network is being coordinated on an entirely voluntary basis and is looking for further labs to join the effort. “We hope existing equipment can be used in situ with qualified staff volunteering to conduct the tests. We are able to provide guidance, protocols, documentation and reporting,” Fischer added.
The Fischer Family Trust has also made £1 million in funding available to support the purchase of consumables for the tests if labs are unable to cover these.
For more information about the Covid-19 Volunteer Testing Network, visit: www.covid19-testing.org
NanoPass shares proprietary MicronJet microneedle to assist in development of a Covid-19 vaccine
, /in Corona News, E-News /by 3wmediaNanoPass is sharing its proprietary MicronJet microneedle device with leading vaccine and immunotherapy companies around the world to assist in development of a Covid-19 vaccine.
The NanoPass device targets immune cells of the skin by harnessing the skin’s potent immune system to improve vaccines and/or to dramatically reduce the dose while achieving the same immunity.
“The human skin is our first layer of defence against many infectious diseases,” says Yotam Levin, MD, CEO of NanoPass. “The skin contains specialized Dendritic Cells that process and induce strong immune responses – that’s why microneedle injections enable reduction of vaccine doses by five-fold, thereby reducing overall cost, required capacity and production time. We believe a reliable injection into the skin is critical for successful activation of broad and effective immune responses, which should be explored for most injectable vaccines.”
The company’s technology is supported by more than 55 completed/ongoing clinical studies with various vaccines and vaccine platforms, including H1N1, H5N1 and live attenuated VZV vaccine, that have shown improved immunogenicity and significant dose-sparing. Pre-clinical evidence with mRNA and DNA vaccines showed promising results.
NanoPass has previously supported US CDC in a Phase 3 infant polio vaccination trial; with ITRC on PPD skin testing; in Type 1 Diabetes immunotherapy; and supported NIAID with devices to evaluate immunogenicity of a pandemic flu vaccine; and multiple vaccine pharma.
NanoPass Technologies flagship product, the 0.6 mm MicronJet, is the first true (<1 mm) microneedle to receive FDA clearance as an intradermal delivery device for substances approved for delivery below the surface of the skin. It is supported by extensive clinical data and regulatory approvals in most major markets including the US, Europe, China and Korea.
IVD assay iAMP Covid-19 Detection Kit receives CE Mark
, /in Corona News, E-News /by 3wmediaFujirebio Europe has received the CE mark for the molecular IVD assay iAMP Covid-19 Detection Kit from its partner Atila Biosystems. The qualitative detection kit is based on real-time fluorescent reverse transcription isothermal amplification, eliminating the need for RNA extraction.
The detection kit was also granted Emergency Use Authorization by the US Food and Drug Administration on April 10.
The iAMP COVID-19 Detection Kit can be run on a Real-Time PCR PowerGene 9600 Plus or any other qPCR automate capable of measuring fluorescence in FAM/HEX channel in real-time.
The new iAMP COVID-19 molecular assay complements the existing panel of biomarkers available on the LUMIPULSE® G System for infection (PCT, Ferritin), inflammation (IL-6) and epithelial lung injury (KL-6) to predict disease severity in patients infected with SARS-CoV-2.
Products from Atila Biosystems are available through Fujirebio’s European affiliates and through a large portion of Fujirebio’s existing or new European distribution network.
For more information, visit: www.fujirebio.com/en/contact
BIOHIT supplies COVID-19 detection kits in the UK
, /in Corona News, E-News /by 3wmediaBIOHIT Healthcare is distributing test kits for the diagnosis of both current and past COVID-19 infections to help in the fight against coronavirus in the UK. The new product line includes the MutaPLEX® Coronavirus kit from Immundiagnostik AG (IDK) – a real-time RT-PCR assay to screen for infected individuals – and Epitope Diagnostics Inc’s (EDI’s) immunodiagnostic tests for IgM and IgG COVID-19 antibodies, to detect past infections.
The IDK MutaPLEX coronavirus screening assay allows the detection of SARS-CoV-2 viral RNA in a variety of biological specimens, especially nasal/throat swabs. This real time RT-PCR kit contains all the reagents, primers and dual-labelled probes required for the amplification and simultaneous differentiation of RNA from SARS-CoV-2 and other betacoronaviruses, as well as house-keeping genes designed to prevent false negative results due to insufficient sample collection or transport problems.
EDI’s Novel Coronavirus COVID-19 ELISA kits provide qualitative detection of antibodies in patient serum, indicating a past COVID-19 infection. The IgM assay provides the earliest immunodiagnostic indication of an infection, while the IgG test can be used to aid detection and provide an indication of long-term immunological response, making it particularly useful in cases where clustering is suspected or differential diagnosis is required.
These tests extend and complement BIOHIT’s repertoire of diagnostic kits for gastroenterology, aiding the evaluation of patients with both GI and upper respiratory complaints, as COVID-19 may include stomach and bowel symptoms in some cases. Inflammatory bowel disease patients being treated with immunosuppressive agents should also be considered at high risk for COVID-19, making differential diagnosis essential.
Avacta Group collaborates with Adeptrix to develop a high throughput Covid -19 antigen test
, /in Corona News, E-News /by 3wmediaUK-based Avacta Group, the developer of Affimer biotherapeutics and reagents, will collaborate with US-based Adeptrix to develop a high throughput Covid -19 antigen test using Adeptrix’s proprietary bead-assisted mass spectrometry (BAMS) platform.
The Affimer-based BAMS coronavirus antigen test that will provide clinicians with a significant expansion of the available testing capacity for Covid-19 infection in hospitals.
Adeptrix’s novel BAMS platform combines enrichment of the sample to improve sensitivity with the power of mass-spectrometry to improve specificity. Hundreds of samples per day can be analysed by a single technician using BAMS, exceeding the capacity of single PCR machine, making BAMS a very attractive high throughput technique for Covid-19 screening in the clinical setting.
The diagnostic test will allow hospitals around the world to utilise their existing installed base of mass spectrometers that are not currently used for Covid-19 testing, thus contributing significantly to the increase in global testing capacity. Avacta’s recently developed Affimer reagents that bind the SARS-COV-2 spike protein will be used to provide the capture and enrichment of the virus particle from the sample which could be saliva, nasopharyngeal swabs or serum.
The companies are aiming to have a BAMS test ready for clinical validation, regulatory approval and manufacturing in June. Adeptrix and Avacta are already in discussion with large-scale manufacturing partners to rapidly deploy this new high throughput test.
Dr Alastair Smith, Chief Executive Officer of Avacta Group, commented: “We believe that the BAMS test will be hugely attractive as an adjunct to PCR testing because it uses laboratory equipment that is already in hospital labs but not currently used for Covid-19 testing so it provides incremental testing capacity.
I have made it clear that we intend to partner the SARS-COV-2 spike protein Affimer reagents with several select companies to support antigen test development on multiple diagnostic test platforms. This will contribute most effectively to the urgent need to increase antigen testing capacity globally and maximise the commercial return to Avacta. Adeptrix is one example of this and other discussions are underway. I look forward very much to further updating the market in the near future.”
Dr. Jeffrey C. Silva, Director of Product Development, Adeptrix Corporation commented: “Mass spectrometry can enhance the diagnostic utility of immunoassays, as it is capable of monitoring both existing and emerging viral strains by accurately measuring the molecular components of the virus. BAMS provides an ideal multiplexing platform to obtain higher specificity for monitoring Covid-19 infection.”
DefiniGEN launches tool to support in vitro intestinal research of Covid-19
, /in Corona News, E-News /by 3wmediaUK-based iPSC (induced pluripotent stem cells) disease modelling company DefiniGEN has identified iPSC-derived intestinal organoids that could be used to help structure in vitro studies of the biology of SARS-CoV-2 infection across cohorts of multiple patients.
While SARS-CoV-2 primarily targets the respiratory system, studies have shown that it also infects and multiplies within the intestinal epithelium. IPSC-derived organoids exhibit characteristics that closely mimic the in vivo intestinal epithelium, making them a valuable surrogate model for studying the virus.
The company says their iPSC-derived intestinal organoids provide a unique in vitro system to model the human intestine. The organoids display a polarized epithelium and harbour a mixture of cell types normally present in the primary intestinal epithelium barrier in vivo, including goblet cells, Paneth cells, enterocytes, LRG5+ stem cells, and enteroendocrine cells. The organoids polarise, form crypt structures and grow villi at the apical surface, and are shown to secrete mucus in a similar manner to primary human gut tissue.
DefiniGEN points out that several studies have proven that angio-tensin-converting enzyme 2 (ACE2) expression in host cells is required for SARS-CoV-2 recognition and infection. Activity of membrane proteases such as TMPRSS2 cleaves the coronavirus’ Spike protein and facilitates the membrane fusion with the host cell. Human intestine is one of the few human tissues with high expression of both ACE2 and TMPRSS2 therefore is a good candidate to study Covid-19 and the mechanisms of the SARS-CoV-2 infection.
Additionally, DefiniGEN have a platform to generate various patient-derived intestinal models which could support population studies, using many different donors with diverse ethnic profiles.
Such studies are useful as there is growing evidence that ethnic differences are a major factor in patients showing a severe response to Covid-19.
DefiniGEN’s differentiation platform is optimized to enable successful generation of intestinal organoids from a diverse range of patients. Patient skin fibroblasts or PBMCs can first be reprogrammed to iPSC, and then differentiated to produce mature intestinal organoids which carry the original patient genetics, and so manifest a gut model specific to that donor.
For more information, visit www.definigen.com/products/intestinal/covid-19
Siemens starts worldwide shipping of total antibody test for COVID-19
, /in Corona News, E-News /by 3wmediaSiemens Healthineers announced late May that it is now shipping worldwide its laboratory-based total antibody test to detect the presence of SARS-CoV-2 IgM and IgG antibodies in blood. The test received the CE mark and data has demonstrated 100 percent sensitivity and 99.8 percent specificity. The total antibody test allows for identification of patients who have developed an adaptive immune response, which indicates recent infection or prior exposure.
The US FDA has issued an Emergency Use Authorization (EUA) for its laboratory-based total antibody test.
Siemens says it is prepared to ramp up production as the pandemic evolves with capacity exceeding 50 million tests per month across its platforms starting in June.
The antibody test is now available on the largest installed base in the U.S. and one of the largest in the world with 20,000 Siemens Healthineers systems installed worldwide. This includes the Atellica Solution immunoassay analyser, which can run up to 440 tests per hour and enables a result in just 10 minutes. By detecting both IgM and IgG antibodies, the test provides a clearer clinical picture over a longer period of time as the disease progresses.
The antibody test also is available on the company’s installed base of ADVIA Centaur XP and XPT analysers, which can test up to 240 samples per hour, with a result in 18 minutes.
Importantly, the test detects antibodies to a key spike protein on the surface of the SARS-CoV-2 virus, which binds the virus to cells with a distinct human receptor found in lungs, heart, multiple organs and blood vessels. Studies indicate that certain (neutralizing) antibodies to the spike protein can disarm SARS-CoV-2, presumably by interfering with the ability of the virus to bind, penetrate and infect human cells. Multiple potential vaccines in development for SARS-CoV-2 include the spike protein within their focus.
Avacta ships SARS-COV-2 Affimer reagents to Cytiva and Adeptrix for diagnostic test development
, /in Corona News, E-News /by 3wmediaAvacta Group plc, the developer of Affimer biotherapeutics and reagents, has started shipping Affimer reagents for COVID-19 antigen testing to its diagnostic test development partners.
The Group recently reported that it had generated multiple, highly specific Affimer reagents that bind the SARS-COV-2 viral antigen and do not cross-react with SARS, MERS and other closely related coronaviruses. These Affimer reagents will be used to develop a point-of-care saliva based COVID-19 antigen test strip by Cytiva (formerly GE Healthcare Life Sciences) for CE marking in Europe and FDA approval in the United States.
The Affimer reagents have been manufactured by Avacta in the quantities required for test development and are being sent to Cytiva. The reagents are also being provided to Adeptrix with whom Avacta has announced that it will develop a COVID-19 laboratory test to run on hospital mass spectrometers using Adeptrix’s proprietary BAMS assay platform.
The Affimer reagents have been studied further by Avacta and this has shown that there are Affimer reagents that can work in pairs, both binding to the spike protein at the same time. This allows tests to be developed that detect both the intact virus particle and the detached spike proteins which become separated from the virus particle during the development of the COVID-19 disease, which may also be important in monitoring disease progression.
Cytiva and Avacta will now work to develop rapid test strips for the detached spike protein and for the intact virus particle. Adeptrix is working to develop a prototype BAMS test. Both of these tests will indicate whether a person has the infection at that moment.