WHO/BS/2016.2298 Rev.1 ENGLISH ONLY EXPERT COMMITTEE …
Transcript of WHO/BS/2016.2298 Rev.1 ENGLISH ONLY EXPERT COMMITTEE …
WHO/BS/2016.2298 Rev.1
ENGLISH ONLY
EXPERT COMMITTEE ON BIOLOGICAL STANDARDIZATION
Geneva, 17 to 21 October 2016
Requests to initiate new WHO reference material projects for in vitro diagnostic
devices and reagents
Document prepared by the WHO Secretariat, based on inputs from WHO Collaborating Centres
supporting biological standardization activities. Note:
This document has been prepared for the purpose of inviting comments and suggestions on the
proposals contained therein, each of which will be considered by the Expert Committee on
Biological Standardization. The proposals have not yet been endorsed by the Expert Committee.
Comments on the proposals MUST be received by 3 October 2016 and should be addressed to
the World Health Organization, 1211 Geneva 27, Switzerland, attention: Technologies, Standards
and Norms (TSN). Comments may also be submitted electronically to the Responsible Officer:
Dr David Wood ([email protected]). The outcome of the deliberations of the Expert Committee
will be published in the WHO Technical Report Series.
© World Health Organization 2016
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Introduction
The provision of global measurement standards is an important normative activity of WHO.
Biological reference preparations that are accepted internationally enable the efficacy, quality, purity
and safety of very many biological medicines, used in the prevention, treatment or diagnosis of
disease or conditions, to be stated in a common language worldwide. International biological
reference standards support the use of many biological and immunological assays for the quality
control of a wide range of biologicals including therapeutics, blood-derived products, vaccines and
immunological products of traditional types as well as those derived from modern biotechnological
approaches. They also have important applications in the standardization of materials and
approaches used in medical diagnostics such as diagnosing disease, monitoring therapy, blood safety,
and public health applications (e.g. monitoring immune status, screening for disease or susceptibility)
or otherwise characterizing biological material from individuals.
WHO biological reference standards are widely used in the development, evaluation, standardization
and control of products by industry; by regulatory authorities; and also in biological research in
academia and scientific organizations. They play a vital role in facilitating the transfer of laboratory
science into worldwide clinical practice and the development of safe and effective biologicals.
The timely development of new reference materials and standards is critically important to harness
scientific developments for new biologicals. At the same time, the active management of the
existing inventory of reference preparations requires a carefully planned programme of work to
replace established materials before the stock of containers, which comprises the standard, is
exhausted.
Considerations for assignment of priorities to development of WHO International Biological
Measurement Standards or Reference Reagents have been published (WHO TRS 932, Annex 2,
Appendix 1, 2005). These considerations are used as guiding principles by the Secretariat and the
WHO Collaborating Centres to develop a proposed programme of future work. To facilitate and to
improve transparency in the priority setting process, a simple tool has been developed which
describes the salient features of each new project proposal.
This document provides a means for the Committee and other stakeholders to review and comment
on new proposals that are under consideration. The proposals in this document
(WHO/BS/2016.2298) cover requests to initiate new projects in the areas of in vitro diagnostic
devices and reagents (Appendix 1).
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Appendix 1 In vitro diagnostic devices and reagents
Proposed new projects
1) BRAF V600E (proposed WHO 1st International Genomic Reference Panel) (page 4)
2) Human epidermal growth factor receptor 2 (HER2, official gene symbol ERBB2) (proposed
WHO 1st International Genomic Reference Panel) (page 6)
3) Syphilitic plasma IgG and IgM (proposed WHO 2nd International Standard) (page 8)
4) Syphilitic plasma IgG (proposed WHO 2nd International Standard) (page 8)
5) Respiratory Syncytial Virus (RSV) RNA for NAT (proposed 1st WHO International
Standard) (page 9)
6) Influenza virus types A and B RNA for NAT (proposed 1st WHO International Standard)
(page 11)
7) Anti-Zika virus antibody (IgG/IgM) (proposed WHO 1st International Standard) (page 13)
8) Anti-Chikungunya virus antibody (IgG/IgM) (proposed WHO 1st International Standard)
(page 15)
9) Ebola virus antigen (proposed 1st International Reference Reagent) (page 18)
10) Human Immunodeficiency Virus type 2 (proposed 2nd International Standard for HIV-2
NAT) (page 20)
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Proposal (title) Proposed 1st WHO International Genomic Reference Panel for BRAF V600E
Proposer (name of Institution)
NIBSC Principal contact Jennifer Boyle
Rationale BRAF mutations are present in approximately 37-50% of malignant melanomas (plus many other solid tumours including colorectal cancer, papillary thyroid carcinoma and non-small-cell lung carcinoma). Activating mutations induce stimulation of the MAP kinase pathway with subsequently increased cellular proliferation and survival. Of all BRAF mutations, 80-90% are BRAF V600E.
The BRAF V600 mutated protein is an obvious candidate for drug-targeting with inhibitor drugs in ongoing development; vemurafenib and dabrafenib are approved for metastatic melanoma treatment in patients with BRAF V600 mutations, with BRAF V600 molecular genotyping mandatory.
It is therefore important to standardise the molecular diagnosis of BRAF V600E-positive patients and monitor treatment response by quantification of BRAF V600E by the provision of a WHO international genomic reference panel to enable accurate and sensitive diagnostic testing.
Anticipated uses and users
The panel would be used by manufacturers for the calibration of diagnostic kits and by clinical & reference laboratories in the calibration of secondary standards used in multiple routine diagnostic assays for BRAF V600E detection.
Standardisation could be applied to assays utilizing both traditional formalin-fixed, paraffin-embedded tissue and ‘liquid biopsy’ circulating tumour DNA, the latter requiring highly sensitive detection and timely standardisation as a rapidly emerging technology.
Source/type of materials Two purified genomic DNAs extracted from cell lines; one BRAF wild-type, one BRAF V600E. Freeze-dried at approximately 2500 ampoules per material.
Outline of proposed collaborative study
The collaborative study would evaluate the materials using a variety of diagnostic genotyping techniques. Quantitative data derived from the collaborative study would be used to establish a consensus value for each of the materials.
Issues raised by the proposal
Feedback from potential end-users and resource availability will determine if the panel should be provided at a range of BRAF V600E levels as ‘ready-to-use’ standards, or as a single BRAF V600E standard for end-user’s dilution (along with the BRAF wild-type material).
V600E is the predominant BRAF mutation in metastatic melanoma; other, much rarer mutations would not be covered by the panel (V600E complex (V600Ec), V600D, V600K and V600R).
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Action required ECBS to endorse proposal
Proposer's project reference
TDI-00006 Date proposed: 1st July 2016
CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)
Approval status of medicine or in vitro diagnostic method
The approval of vemurafenib and dabrafinib, two BRAF inhibitor drugs, have made BRAF V600 molecular genotyping mandatory in selecting patients who will benefit from these therapies. Companion diagnostic assays have been approved for both drugs, but their use is not obligatory. The high cost of these genotyping systems means that a variety of unregulated tests of variable quality continue to be used.
Number of products or methods
Materials to enable standardisation of all commonly used PCR-based assays for BRAF V600, as well as next-generation sequencing.
Public health importance
The incidence and mortality rates of melanoma have risen sharply throughout the world over the past few decades. With over 130,000 new case of melanoma diagnosed globally each year, there is a strong need to improve the quality of diagnosis and treatment.
Global importance
Melanoma occurs in all countries, with Caucasians living in low and middle latitude countries at the highest risk.
Global need from regulatory & scientific considerations
Accurate genotyping is required to ensure patients are matched to the optimal drug treatment. Tumour biopsies from which DNA is extracted contain variable levels of tumour cells. Even commercial assays have been demonstrated to lack sufficient analytical sensitivity to detect mutations in small tumours, unless macrodissections are performed before DNA is extracted. Without sensitivity standards and calibrants, satisfactory assay performance cannot be guaranteed.
An international reference panel will enable global standardisation of BRAF V600 genomic diagnostics to ensure a consistent, accurate and safe approach.
ECBS outcome [BLANK]
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Proposal (title) Proposed 1st WHO International Genomic Reference Panel for HER2
Proposer (name of Institution)
NIBSC Principal contact Jennifer Boyle
Rationale Human epidermal growth factor receptor 2 (HER2, official gene symbol ERBB2) is a member of the epidermal growth factor receptor family of receptor tyrosine kinases. ERBB2 is amplified (and thus HER2 protein is overexpressed) in 18-20% breast cancers, and is associated with increased tumour aggressiveness. HER2 overexpression is also associated with more aggressive forms of ovarian, stomach and uterine cancer.
Detection of HER2 overexpression is clinically actionable, with HER2-positive tumours treatable with herceptin (trastuzumab). Nucleic-acid based ERBB2 quantification methods are of increasing diagnostic use (particularly RNA). It is therefore important to facilitate timely standardisation of the molecular diagnosis of HER2 overexpression-positive patients and monitoring of treatment response by quantification of ERBB2 by the provision of a WHO international genomic reference panel to enable accurate and sensitive diagnostic testing.
Anticipated uses and users
The panel would be used by manufacturers for the calibration of diagnostic kits and by clinical and reference laboratories in the calibration of secondary standards used in multiple routine diagnostic assays for ERBB2 characterisation.
Standardisation could be applied to assays utilising both formalin-fixed, paraffin-embedded tissue and fresh-frozen solid tumour biopsies.
Source/type of materials The panel would comprise freeze-dried cells for DNA or RNA extraction at approximately 2500 ampoules per material.
One cell line with confirmed both high ERBB2 expression and copy number would be chosen from several in-place candidates. Similarly, ERBB2 status of a likely wild-type cell line would be confirmed. A dilution series of the high ERBB2 mRNA expression and copy number cell line in the ERBB2 wild-type cell line would generate four materials providing low, low medium, high medium and high values for both mRNA expression and DNA copy number.
Outline of proposed collaborative study
The collaborative study would evaluate the materials using a variety of diagnostic genotyping techniques. Quantitative data derived from the collaborative study would be used to establish a consensus value for each of the materials.
Issues raised by the proposal
The National Institute for Standards and Technology (NIST) has produced a standard reference material for ERBB2. However, this is of genomic DNA suitable only for the measurement of DNA copy number, and not mRNA expression which is much more commonly used in diagnostics.
Action required ECBS to endorse proposal
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Proposer's project reference
TDI-00027 Date proposed: 1st July 2016
CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)
Approval status of medicine or in vitro diagnostic method
Herceptin (trastuzumab) is approved for breast cancer treatment in patients with HER2 overexpression. The European patent expired in 2014, and with the USA patent expiring in 2019, biosimilars are already emerging.
Laboratory diagnostics by immunohistochemistry for HER2 protein overexpression and fluorescence in situ hybridization for ERBB2 gene amplification are known to be inaccurate; nucleic-acid based quantification methods are of increasing use (particularly for mRNA).
Number of products or methods
Materials to enable standardisation of all commonly used DNA- and RNA-based quantitative PCR-based assays for ERBB2.
Public health importance
There are 1.6 million new breast cancer cases globally each year, of which approximately 20% are HER2 overexpression-positive.
Herceptin biosimilars may vary in quality, efficacy and safety. Provision of an international reference panel to calibrate diagnostic assays will ensure biosimilar products are safe and effective.
Global importance
Just over half of breast cancer cases (including HER2 overexpression-positive cases) are from less developed countries (data from World Cancer Research Fund International).
Global need from regulatory & scientific considerations
Accurate genotyping is required to ensure patients are matched to the optimal drug treatment. The release of Herceptin biosimilars will bring effective treatments to many more patients than previously.
No standards currently exist for the measurement of RNA-based HER2/ERBB2 over-expression assays, an increasingly typical diagnostic approach.
An international reference panel will enable global standardisation of HER2/ERBB2 genomic diagnostics to ensure a consistent, accurate and safe approach.
ECBS outcome [BLANK]
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Proposal (title) The second International Standard for Syphilitic plasma IgG and IgM & second International Standard for Syphilitic plasma IgG
Proposer (name of Institution)
e.g. NIBSC Principal contact David Padley
Rationale Based on current uptake the Syphilitic plasma IgG and IgM (human)(1st International Standard) 05/132 & the Syphilitic plasma IgG (human)(1st International Standard) 05/122 will run out in 2-3 years
Anticipated uses and users
Reference labs, hospital labs, GUM/STI clinics, manufacturers
Source/type of materials Syphilitic human plasma
Outline of proposed collaborative study
Collaborative study with candidate samples freeze-dried, frozen bulks and clinical samples for commutability. 15- 20 laboratories hopefully participating.
Issues raised by the proposal
None.
Action required ECBS to endorse proposal
Proposer's project reference
Date proposed: 05-07-2016
CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)
Approval status of medicine or in vitro diagnostic method
Number of products or methods
Public health importance
Global importance
Global need from regulatory & scientific considerations
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Proposal (title) 1st WHO International Standard for Respiratory Syncytial Virus (RSV) RNA
for NAT
Proposer (name of Institution)
NIBSC Principal contact Clare Morris
Rationale Respiratory tract infections can be caused by a plethora of pathogens, all with varying degrees of severity. RSV is a seasonal infection during winter months and is major cause of lower respiratory tract infections often causing severe disease in the elderly or very young; in many cases hospitalisation is required. Early diagnosis is therefore critical for correct diagnosis and therefore early application of treatment. There is no vaccine for RSV.
Laboratories are moving away from serological screening towards more sensitive molecular methods. However data returned through EQA schemes indicates that there is a large degree of variation across all assays. Assays are typically reporting in qualitative format, however the large variation in results reported on the same sample suggests the incidence of incorrect reporting both as false negative and false positive occurs. Without the availability of an international standard it is also very difficult to accurately determine LOD and LOQ. RSV can be divided into two main subtypes, A and B. It is unknown whether current molecular assays detect both strains with equivalent sensitivity
Anticipated uses and users
Manufactures of RSV assays
Laboratories with in house assays
Providers of secondary reference materials
Source/type of materials Literature reports indicate that RSV can be grown to high titre in HEp-2 and MA-160 cells in tissue culture and therefore it is anticipated that candidate material for RSV A and RSV B will be produced in this way.
Laboratories will typically screen for RSV in a range of respiratory samples – sputum, throat swabs, BAL. In collaboration with clinical laboratories, such samples will be sought for inclusion in the study to contribute towards commutability assessment.
Outline of proposed collaborative study
Study to include up to 30 international laboratories running a variety of commercial and in house assays
Issues raised by the proposal
Availability of clinical material to assess commutability. Availability of clinical material to assess commutability may be limited. Typically a high volume (>500ml) of individual patient material is needed to aliquot amongst study participants in order for identical un manipulated clinical material to be assessed as would mimic the situation in diagnostic laboratories.
Action required ECBS to endorse proposal
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Proposer's project reference
Date proposed: 17 June 2016
CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)
Approval status of medicine or in vitro diagnostic method
Diagnostic methods are CE marked where they are commercial, however status of in house methods is unknown
Number of products or methods
4 commercial assays known for single detection of RSV, multiple assays >6 for the multiplex detection of RSV. Unknown number of in house methods
Public health importance
PHE figures indicate that typically 1000-1200 cases are reported in the UK during the winter season.
Global importance
WHO data indicates that RSV is responsible for in the order of 30 million global respiratory tract infections annually
Global need from regulatory & scientific considerations
Lack of vaccine material and the high number of hopitailsations resulting from RSV infections in the young and elderly lead to an urgent need for accurate reporting.
ECBS outcome [BLANK]
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Proposal (title) 1st WHO International Standard for Influenza types A and B for NAT
Proposer (name of Institution)
NIBSC Principal contact Clare Morris
Rationale Respiratory tract infections can be caused by a plethora of pathogens, all with varying degrees of severity. Influenza is a major cause of respiratory tract infections. Influenza has many subtypes determined by the combination
of hemagglutinin (HA) and neuraminidase (NA), two proteins on the surface of
the virus: The severity of disease can be dependent on the combination of HA and NA. Influenza has been devised into 3 types, A, B and C. Types A and B are known to be capable of causing the most severe form of illness. Assays are reported to be capable of detecting both type A and B with equivalent sensitivity. However data returned through EQA schemes indicates that there is a large degree of variation across all assays, it is unknown to what degree this is caused by variant or type. The use of molecular assays is becoming increasing more frequently in the determination of a respiratory illness being caused by influenza. In particular the use of rapid assays.
Anticipated uses and users
Manufactures of Influenza A/B assays
Laboratories with in house assays
Providers of secondary reference materials
Source/type of materials Literature reports indicate that both Influenza A and B can be grown to high titre in tissue culture and therefore it is anticipated that candidate material for A and B will be produced in this way.
The selection of appropriate strain is addressed below in issues raised.
Laboratories will typically screen for influenza in a range of respiratory samples – sputum, throat swabs, BAL. In collaboration with clinical laboratories, such samples will be sought for inclusion in the study to contribute towards commutability assessment.
Outline of proposed collaborative study
Study to include up to 30 international laboratories running a variety of commercial and in house assays
Issues raised by the proposal
Availability of clinical material to assess commutability may be limited. Typically a high volume (>500ml) of individual patient material is needed to aliquot amongst study participants in order for identical un manipulated clinical material to be assessed as would mimic the situation in diagnostic laboratories.
Selection of an appropriate strain that remains representative over time. It would be appropriate to evaluate a panel of materials and this may need to take to form of a pilot study to identify, if possible, 2/3 strains to put forward into the main study. Additional guidance needs to be sought from experts in this field. Currently strains to which assays commonly refer to as detecting are H1N1 and H3N2, these also feature widely in UK and WHO incidence reports. These viral strains are also reflected in many run control reagents.
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As a result these would be a good inclusion in the panel. It. Recombination events within the virus lead to frequent changes in the HA and NA combinations. Whilst commercial assays often target conserved areas of the influenza genome, it would be wise to assess the validity of the standard on an annual basis. This could be achieved by inclusion in an EQA scheme as a means of reassessing performance with a range of assays.
Action required ECBS to endorse proposal
Proposer's project reference
Flu A/B 2016 Date proposed: 17 June 2016
CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)
Approval status of medicine or in vitro diagnostic method
Diagnostic methods are CE marked where they are commercial, however status of in house methods is unknown
Number of products or methods
Commercial assays are available for single detection of Influenza A/B however many assays(>6) are on the market for the multiplex detection of Flu A/B these often fall into the rapid test field. Unknown number of in house methods
Public health importance
PHE figures indicate that typically 150,000 cases per week reported in the UK during the winter season.
Global importance
WHO data indicates that Flu A/B infections are responsible for 3 to 5 million cases of severe illness, and about 250 000 to 500 000 deaths annually
Global need from regulatory & scientific considerations
A vaccine is available however not adopted by all
ECBS outcome [BLANK]
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Proposal (title) The 1st International Standard for antibody (IgG/IgM) to Zika virus
Proposer (name of Institution)
NIBSC Principal contact Mark Page
Rationale In support of the WHO response to Zika virus disease, NIBSC has been requested to lead in the development of an International Standard for use in calibration and control of Zika antibody assays. Accurate diagnosis of Zika virus infection is critical for subsequent healthcare decisions particularly for pregnant women. The virus has a short viraemia and PCR based tests are only useful during this period (~9 days) after onset of symptoms (rash, fever). Diagnosis is then carried out by serology tests to measure the antibody response to the virus. Current tests are prone to high cross reactivity against other arboviruses (Dengue in particular) that are spread by the same mosquito vector. Therefore it is critical that accurate diagnosis is available and standardization of the tests is required to improve sensitivity and specificity measurements. Both IgM and IgG are currently measured to determine prior exposure where IgM would indicate a recent infection that is used in secondary testing after a negative PCR result.
Anticipated uses and users
Primary intended use is for calibration of assays used to measure Zika antibody IgM and IgG levels in human serum by ELISA and neutralization assays. Recommendation for this purpose will be subject to satisfactory demonstration of commutability. Primary users will be public health and other clinical laboratories, kit manufacturers and Zika vaccine manufacturers (for clinical trials).
Source/type of materials Human serum/plasma from subjects recently infected with Zika virus.
Serum from immunized transchromosomal bovines that produce fully humanized IgG.
Outline of proposed collaborative study
assess the suitability of different antibody preparations to serve as the Intenational standard with an assigned unitage per mL for use in the harmonization of Zika serology assays. There is no international conventional reference measurement procedure for Zika virus antibodies and the unitage will not be traceable to the International System of Units (SI) of quantity.
characterise the antibody preparations in terms of reactivity/specificity in different assay systems.
assess each preparation’s potency i.e. readout in a range of typical assays performed in different laboratories for both IgG and IgM classes
assess commutability i.e. to establish the extent to which each preparation is suitable to serve as an interim standard for the variety of different samples and assay types.
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Issues raised by the proposal
Material transfer agreements and sourcing material from embargoed countries may add to the timeline.
Action required e.g. ECBS to endorse proposal
Proposer's project reference
Date proposed:
CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)
Approval status of medicine or in vitro diagnostic method
Number of products or methods
Experimental ELISA kits in development. Commercial ELISA kits are being rolled out. Neutralisation assay measured by plaque reduction.
Public health importance
Increasing numbers of countries are reporting cases. Zika association with microcephaly and Guillain-Barre syndrome is of high importance with potential for global impact. WHO have declared the outbreak as a Public Health Emergency of International Concern.
Global importance
As above
Global need from regulatory & scientific considerations
Standardised and calibrated assays are vital for accurate diagnosis of Zika (see above)
ECBS outcome ECBS to endorse
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Proposal (title) Chikungunya virus reference reagent for serology
Proposer (name of
Institution)
PEI Principal
contact
Sally Baylis
Rationale The mosquito-borne Chikungunya virus (CHIKV) is a
member of the Alphavirus genus in the Togaviridae family.
Chikungunya was first identified in Tanzania in the early
1950s. The disease occurs not only in Africa but also in
Asia and the Indian subcontinent and, since 2013, has
spread to the Americas, particularly central and Southern
areas. Small outbreaks have also occurred recently in
Europe.
The diagnosis of Chikungunya requires a variety of tests
including detection of IgM and IgG antibodies. Co-
circulation of Chikungunya virus with Dengue virus (DENV)
and Zika virus (ZIKV) frequently occurs and infections
cause by these viruses share common signs and symptoms
in infected patients. Accurate diagnosis and discrimination
of CHIKV from other virus infections is important for patient
care. Analysis of IgM antibodies is particularly useful for the
confirmation of acute infection. Anti-CHIKV IgG may also be
detectable during acute infection. Anti-CHIKV IgG is also a
marker of past CHIKV infection and seroprevalence and
anti-CHIKV and indication previous exposure of different
populations to CHIKV infection.
Anti-CHIK assays vary in their performance (for example,
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Niedrig et al. Clin Microbiol Infect. 2009;15:880 and Prat et
al. Emerg Infect Dis. 2014;20:2129).
Anticipated uses
and users
Clinical laboratories, particularly arbovirus reference
laboratories. Research laboratories and organizations
developing CHIKV vaccines. IVD manufacturers.
Source/type of
materials
Sera to be sourced through a national and international
collaborations.
Alphavirus cross reactivity.
Outline of
proposed
collaborative study
Evaluate sera from CHIKV infected patients and potentially
blood donors. These samples will include IgM as well as
IgG reactive sera to distinguish between recent and past
infections.
Issues raised by
the proposal
Action required ECBS to endorse proposal
Proposer's project
reference
Date
proposed:
July 2016
CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)
Approval status of
medicine or in
vitro diagnostic
A number of serological kits are available for the detection
of anti-CHIKV IgM and IgG.
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method
Number of
products or
methods
As above.
Public health
importance
To ensure that assays are of adequate sensitivity/specificity
for the determination of anti-CHIK IgM and IgG for
diagnostic purposes and to facilitate a better understanding
of CHIKV epidemiology globally.
Global importance The rapid spread of CHIKV in the last decade and the
similar clinical presentation with viruses such as dengue
and Zika virus require accurate diagnostic testing.
Global need from
regulatory &
scientific
considerations
As above.
ECBS outcome
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Proposed 1st International Reference Reagent for Ebola virus antigen
Proposal (title) Proposed 1st International Reference Reagent for Ebola virus antigen
Proposer (name of Institution)
NIBSC Principal contact Dianna Wilkinson
Rationale In support of the WHO response to the Ebola virus outbreak, NIBSC has been requested to lead in the development of reference reagent for use in control of Ebola virus antigen assays. Accurate and rapid diagnosis of Ebola virus infection is critical for subsequent healthcare decisions. Patients may present with a fever that could be confused with other fever inducing pathogens particularly malaria which is endemic in much of Africa. Rapid diagnostic kits such as those used at the point of care allow confirmation of ongoing virus infection within ~15 minutes from a simple blood prick sample. The read out is a colour reaction on a solid phase stick that gives an easily interpretable yes or no answer. A rapid response is vital to facilitate appropriate healthcare provision particularly the isolation of infected individuals. Other ELISA based assays may also be used in other settings.
Anticipated uses and users
Primary intended use is for a monitor for Ebola antigen assays. Recommendation for this purpose will be subject to satisfactory demonstration of commutability. Primary users will be public health and other clinical laboratories, kit manufacturers.
Source/type of materials Full length, native, VP40 recombinant protein, expressed in E.coli. Based on 2014 Kissidougou – C15 strain
Virus-like particles developed by expression of 3 plasmids for VP40, NP and GP in HEK293T cells. Based on H. sapiens wt/GIB/2014/Kissidougou – C15 Ebola strain. VLPs are purified on 205 sucrose cushion in phosphate buffer
Soluble GP, partial nucleoprotein NP and full length VP40 produced in E.coli using Ebola Mayinga sequences, supplied in carbonate buffer by SD Biosensor Inc., S. Korea
9 coded samples will be entered into the study, formulate in thrombinised and declotted plasma. Samples will be diluted around the detection limit and aliquotted into 120ul and freeze-dried
Outline of proposed collaborative study
assess the suitability of different antigen preparations to serve as the international reference reagent to monitor Ebola antigen assays.
characterise the antigen preparations in terms of reactivity and sensitivity in different assay systems.
assess each preparation’s performance i.e. readout in a range of typical assays performed in different laboratories
assess commutability i.e. to establish the extent to which each preparation is suitable to serve as an interim standard for the variety of different samples and assay types.
Issues raised by the Material transfer agreements and sourcing material from embargoed
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proposal countries may add to the timeline.
Action required ECBS to endorse proposal
Proposer's project reference
TBC Date proposed: 2016
CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)
Approval status of medicine or in vitro diagnostic method
Diagnostic antigen kits are under development, none licensed. Applications for WHO EUAL in progress or are listed.
Number of products or methods
Experimental antigen kits in development. Commercial kits are being rolled out.
Public health importance
Rapid diagnosis of Ebola is vital during an outbreak to identify infected individuals so that they can be treated quickly to prevent death and spread of the virus.
Global importance
As above
Global need from regulatory & scientific considerations
Proper monitoring of kit batches is vital for accurate diagnosis of Ebola (see above)
ECBS outcome
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PROPOSAL FOR A NEW / REPLACEMENT
INTERNATIONAL REFERENCE PREPARATION
Proposal 2nd International Standard for HIV-2 NAT
Proposer NIBSC Principal Contact
Clare Morris
Rationale The 1st WHO IS for HIV-2 RNA was established in 2009 with an assigned value of 1000IU/ml. Many users have contacted NIBSC stating that this value is too low for calibration purposes. It is therefore proposed to formulate a replacement materials at a higher titre (>106). Once established the remaining material for the 1st IS will be withdrawn.
Anticipated uses and users
This standard will used to calibrate HIV-2 NAT assays and secondary HIV-2 NAT standards. It will be used by a range of laboratories including kit manufacturers, blood fractionators, reference laboratories, diagnostics labs, EQA providers and OMCL’s.
Source/type of materials
We propose using the same HIV-2 subtype A strain as was used to manufacture the 1st HIV-2 IS. Live high titre stocks of tissue culture derived virus are already stored under vapour phase liquid nitrogen at NIBSC. Following the successful inactivation of the 1st HIV-2 IS we propose to heat inactivate stock material to formulate the 2nd standard. The viral stocks will be diluted in pooled human plasma prior to lyophilisation.
Outline of proposed collaborative study
A panel of samples would be developed for evaluation in a collaborative study comprising: The current (1st) IS for HIV-2 NAT The proposed candidate 2nd HIV-2 material The frozen liquid bulk material Where possible clinical specimens of HIV-2 positive plasma (however this is expected to be very difficult to obtain) A collaborative study involving 15-20 international laboratories conducting a range of commercial and in-house assays would be invited to participate.
Issues raised by proposal
Possible sourcing of clinical material for commutability assessment
Action required Endorsement of the project
NIBSC Ref VIR-00048 ECBS Endorsement required
WHO/BS/2016.2298 Rev.1
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Approval status of medicine or in vitro diagnostic method
NAT is the most widely used and most sensitive method for the detection of HIV-2 RNA in human serum and plasma.
Number of products or methods
A range of NAT-based methods are available for the detection and quantification of HIV-2 RNA, including both commercial and laboratory-developed assays.
Public health importance
The need to standardise NAT-based assays for HIV-2 is ongoing. NAT is routinely used to manage HIV infections and there remains a major risk of transfusion-transmitted infection due window-period donations.
Global importance
HIV-2 incidence is lower than HIV-1 and typically follows a difference course of infection, with lower viral loads and a higher percentage of longer term non progressors. Not all treatments established for HIV-1 are effective against HIV-2, therefor the accurate and timely diagnosis of HIV-2 infection is critical to ensure correct therapy can be given and to inform the individual of their infection status to prevent the risk of unknown transmissions
Global need from regulatory & scientific considerations
Whilst screening for HIV-2 RNA is not mandated in as many countries as HIV-1, due to the lower incidence and lower titre of an infected individual, accurate and sensitive assays are critical to ensure the safe blood supply in countries where HIV-2 is prevalent.
ECBS outcome [BLANK]