Development and validation of a 3ABC antibody ELISA in Australia for foot and mouth disease.

OBJECTIVE: To measure the diagnostic performance of an Australian-developed ELISA for the detection of antibodies against the non-structural proteins (NSP) 3ABC of the foot and mouth disease (FMD) virus. DESIGN: Test development and validation study. METHODS: The diagnostic specificity was determined using 2535 sera from naïve animals and 1112 sera from vaccinated animals. Diagnostic sensitivity was calculated from the data for 995 sera from experimentally and field-infected animals from FMD-endemic countries in South East Asia. A commercial ELISA detecting antibodies against FMD virus NSP was used as the reference test to establish relative sensitivity and specificity. Bayesian latent class analysis was performed to corroborate results. The diagnostic window and rate of detection were determined at different times using sera from cattle, sheep and pigs before and after infection, and after vaccination and subsequent infection. Repeatability and reproducibility data were established. RESULTS: At 35% test cut-off, the 3ABC ELISA had an overall diagnostic sensitivity of 91.5% and diagnostic specificity of 96.4%. The diagnostic sensitivity in vaccinated and subsequently infected cattle was 68.4% and diagnostic specificity in vaccinated cattle was 98.0%. CONCLUSIONS: The 3ABC ELISA identified field and experimentally infected animals, as well as vaccinated and subsequently infected animals. Diagnostic sensitivity and specificity estimates for other FMD NSP tests are comparable with the results obtained in this study. This NSP ELISA was found to be 'fit for purpose' as a screening assay at the herd level to detect viral infection and also to substantiate absence of infection.

A modified liquid phase (LP) blocking ELISA used to assess type O foot-and-mouth disease virus antigenic variation in Thailand.

A selection of type O foot-and-mouth disease (FMD) viruses isolated in Thailand between 1986 and 1989 were compared to the reference viruses O1 Thailand 1960 (O BKK/60) and O Nakorn Pathom 1965 (O NPT/65) using a liquid-phase blocking ELISA (LP ELISA) to derive serum titres and associated r values. Interpolation techniques were used to increase the precision for estimation of r values through a more accurate estimation of serum titres at predicted equivalent levels of antigen input. Mean r values were 0.45 (for 56 field viruses) relative to O BKK/60 reference virus and 0.56 (for 51 field viruses) relative to O NPT/65. While only two viruses showed considerable difference (r < 0.20) to a reference virus (O BKK/60), 41% and 31% gave r values less than 0.4 for O BKK/60 and O NPT/65 respectively. This indicated antigenic differences between reference and field viruses which may result in a reduction in vaccine efficacy.

Establishment of a typing enzyme-linked immunosorbent assay for foot and mouth disease antigen, using reagents against viruses endemic in Thailand.

Antisera were produced at a central laboratory in Thailand against the endemic serotypes (O, A and Asia 1) of foot and mouth disease (FMD) virus. At a regional veterinary laboratory, these antisera were used in an indirect sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and serotyping of FMD virus (FMDV) antigen. ELISA readings of < 0.10 optical density (OD) units were considered negative. This was verified using fifty tissue samples which were known to be negative for FMDV. The highest mean sample value for three different dilutions was 0.02 OD units. Of a total of 93 samples submitted for antigen typing, 80 (86%) tested positive by ELISA and 13 (14%) were negative. No FMDV was detected in ELISA-negative samples following attempted tissue-culture virus isolation.