An enzyme-linked immunosorbent assay for detection of flavivirus antibodies in chicken sera

Abstract

An enzyme-linked immunosorbent assay (ELISA) was developed to determine the presence of flavivirus antibodies to Murray Valley encephalitis (MVE) and Kunjin viruses in sentinel chicken sera. The development of a quick, reliable assay to detect antibodies to MVE was an essential part of a large-scale surveillance programme to monitor arbovirus activity in Western Australia. This assay was developed for use with alkaline phosphatase conjugated goat anti-mouse IgG using mouse anti-chicken globulin in an intermediate step. There was a significant difference in absorbance values between neutralization test positive and pre-bled sera. However, some sera obtained from sentinel chickens and deemed negative by neutralization demonstrated adsorbance levels above the cutoff level in the ELISA, which reflects the increased sensitivity of this technique. The ELISA test detected antibodies to MVE in chicken sera 7-10 days after infection, whereas these antibodies were only consistently detected by the neutralization test 24 days after infection. Antibodies to both MVE and Kunjin reacted positively with the MVE antigen, but there was little cross-reactivity between this antigen and antibodies to other togaviruses. The main advantages of the ELISA over the neutralization test for detecting antibodies to MVE virus in the sera of sentinel chickens are its greater sensitivity and the speed with which tests can be performed. Results are available within 48 h of receiving specimens and emergency mosquito control measures may then be implemented.