This document outlines the serological assays offered in support of FMD vaccine development and evaluation with specific reference to Eastern Africa. These services are seen as relevant to the developers or evaluators of vaccines intended for use against the four major serotypes of FMD circulating in Eastern Africa (serotypes O, A, SAT1 and SAT2), and are intended to assist those involved in development of vaccines associated with the AgResults Foot and Mouth Disease (FMD) Vaccine Challenge Project.
This project was launched in February 2020, and in the development phase it is expected that vaccine developers will assemble and submit evidence of efficacy of their formulated vaccines against a panel of FMD viruses that are considered representative of those circulating and causing disease outbreaks in Eastern African countries. The level of post-vaccination virus neutralisation titres (VNT) against heterologous viruses is considered an important indicator of likelihood to protect against disease.
The WRLFMD is assembling such a panel of representative viruses, of the 4 serotypes, after consultation process involving the members of the OIE/FAO Reference Laboratory Network, AU-PANVAC, and the Eastern African regional laboratory network (EARLN-FMD). Developers or evaluators of FMD vaccines may request VNT to be performed against the panel of isolates by submitting appropriate bovine vaccine sera to the WRLFMD as set out below (additional testing can also be carried out upon request). Vaccine developers that have sufficient biocontainment facilities may also request to receive the panel of isolates for the purpose of conducting the tests in-house, although for the purpose of the AgResults FMD Vaccine Challenge Project it is understood that the VNT results submitted as part of the application dossier should be from FMD reference laboratories approved for the purpose.
East Africa Panel: guidance for submission of bovine sera for post-vaccination assessment at WRLFMD
- The vaccinal serum should be produced in cattle, which have had no previous exposure to FMD virus through vaccination and/or infection and do not have maternal FMDV antibodies.
- The vaccine used should be the product that will be used in the field (i.e. it should be formulated in the same way, have the same potency and contain the same strains as the final formulated vaccine). NB: It is anticipated that the sera submitted for testing may be polyvalent.
- The vaccination regime adopted should be identical to the schedule recommended to customers for use in field situations. Therefore, a booster vaccination may be given if this is part of the recommended primary course
- Sera should be collected from each animal, at the point of vaccination (day 0) and then at day 21 after vaccination. If a second vaccination is given, then serum should also be collected 10 days later.
- Sera from at least five individual cattle should be submitted. These sera should not be pooled
Testing performed at WRLFMD
- Unless advised, all samples (at all time points) will be tested individually by both virus neutralisation test (VNT) and NSP ELISA.
- For the VNT, the sera will be tested against East African FMD virus isolates representing relevant lineages that are circulating in East Africa.
- All results will be reported on individual sera basis.
- Results will be sent back to the submitter confidentially.
The serum will be evaluated against the following criteria
- Non-structural protein-specific antibodies will be measured using the PrioCHECK FMDV NS Antibody ELISA kit by ThermoFisher Scientific. All pre and post vaccination sera should score negative.
- Virus Neutralisation Titres will be calculated according to the protocol used at WRLFMD (ISO/IEC 17025 accredited SAU-SOP-4). This SOP follows the method described by the OIE Terrestrial Manual 2018, Chapter 3.1.8 – Foot and mouth disease (infection with foot and mouth disease virus). For each isolate used, the strain-specific neutralisation antibody titres will be calculated using the Kaerber equation (Kaerber, 1931), which calculates the neutralisation titre at a virus dose of 100 TCID50.
Kaerber G. (1931). Beitrag zur kollektiven behandlung pharmakologischer reihenversuche. Archive fuer Experimentelle Pathologie Pharmakologie, 162, 480-483. DOI: 10.1007/BF01863914read online
Costs of testing sera raised against vaccines with the FMDV Reference Antigen Panel from WRLFMD, Pirbright Institute, UK.
The following information outlines the costs, provided by WRLFMD, associated with the independent vaccine matching tests that are part of the eligibility requirements for the AgResults FMD Vaccine Challenge Project. As stated in the Competition Rules:
Vaccine testing against the Eastern African FMD panel to be done at an AgResults-approved laboratory, which has demonstrated it has:
- the capability to test against the full approved Eastern African FMD panel according to the agreed methodology,
- no IP / financial conflict of interest with the vaccine development company,
- and accreditation to international standards for the specified testing.
Non-Structural Protein (NSP) Antibody ELISA
£410.00 GBP for up to 40 samples
The cost of one sample or 40 samples is the same.
Virus Neutralisation Test
£45.00 GBP per sample per serotype
Five samples collected at one time point tested against four isolates (1 serotype) the price will be: £45 x 5 samples x 4 isolates = £900
If Manufacturers Vaccinate 5 Animals With Quadrivalent Vaccine
For NSP Testing they will need to supply:
- 5 x day 0 sera for NSP testing
- 5 x day 21 sera for NSP testing
- Total of 10 sera
NSP = £410.00 GBP per 0-40 tests
NSP Cost to submitter = £410.00 GBP ($500.00USD#)
For VNT they will have to supply:
- 5 x day 0 sera for VNT titre analysis against 4 serotypes = 20 sera
- 5 x day 21 sera for VNT titre analysis against 4 serotypes = 20 sera
- Total of 40 sera
VNT= £45 per sample
VNT Cost to Submitter = £1800.00 GBP ($2200.00 USD#)
NSP and VNT - Total Cost to Submitter = £2210.00 GBP ($2720.00 USD#)
# USD subject to exchange rate
Further information about these Reference FMDV antigens (including process to access this panel from WRLFMD) can be found here.