Dr Nilgün Özdural presented a paper (Appendix 27) on the development of a latex agglutination test kit for the detection of A22 Mahmatli and O1 Manisa FMD antigens which is intended for use in the field. The test is highly sensitive and therefore special attention should be given to the time of observation of agglutination. Conclusions •
Once fully validated, real-time, automated RT-PCR could support the ELISA tests for the detection of FMDV in epithelial suspensions and largely remove the necessity for virus isolation in cell culture for the confirmation of secondary cases.
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The comparison of the RT-PCR procedures confirmed the high sensitivity of RT-PCR methods. Nested and novel PCR procedures had a higher sensitivity than conventional RT-PCRs with universal primers, but only quantitative methods scored all samples positive on epithelial suspensions.
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The simplicity and sensitivity of the latex agglutination test (LAT) makes it a good candidate for a pen-side test in endemically infected areas.
Recommendations •
More validation work should be carried out on the automated RT-PCR and the system should be optimised for the testing of probangs and milk.
•
A more extensive comparison of RT-PCR procedures should be performed. Thus, more experimental details such as the position of the assay cut-off, the use of positive and negative controls and details of assay validation need to be made available. Also the specificity of the RT-PCR procedures should be further investigated.
•
The specificity of the LAT kit should be more thoroughly evaluated.
Item 6: Diagnostics - antibody detection Dr Kris De Clercq gave an overview of plans for an EU project for the production of FMD reference sera. The recent outbreaks in Europe and the need to develop and validate new assay systems, including those for the detection of NSP antibodies, have highlighted the importance of developing a panel of such reagents as quickly as possible. The project should involve a consortium of laboratories and an agency specialising in the storage, aliquoting and distribution of such materials. The EU Commission will have to tender the work for competitive bids. Drs David Paton and Bob Armstrong presented results from the FAO Phase XVII serology standardisation exercise (Appendix 28). The reference sera prepared in Phase XV against O Manisa, A22 and C Noville have been adopted by OIE. A new set of reference sera have been prepared against O SKR, A Iran 96 and Asia 1 Shamir. For each strain, a candidate strong, weak and cut-off serum was prepared and distributed, along with a negative serum, to 9 other laboratories. Reagents and a protocol were also supplied for solid phase competition ELISA (SPCE) for sera type O. Results from all the laboratories were presented, including analyses 14
