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A Longitudinal Study of a Novel Dot-Enzyme-Linked Immunosorbent Assay for Detection of Avian Influenza Virus

Lu, Huaguang
Avian diseases 2003 v.47 no.2 pp. 361-369
Influenza A virus, allantoic fluid, bird diseases, chicken eggs, chickens, enzyme-linked immunosorbent assay, epitopes, flocks, hemagglutination, longitudinal studies, monitoring, monoclonal antibodies, selenium, serotypes, specific pathogen-free animals, viruses, Pennsylvania
A monoclonal antibody (MAb)-based dot-enzyme-linked immunosorbent assay (ELISA) has been developed that detected the epitopes specifically associated with avian influenza virus (AIV). The dot-ELISA detected the antigens of AIV directly from clinical and field specimens. Data obtained from experimentally AIV-infected specific-pathogen-free chickens and also the 2001/02 AIV outbreak of serotype H7N2 positive flocks in Pennsylvania indicated that the mean sensitivity (Se) of the dot-ELISA ranged between 45% and 68% and the mean specificity (Sp), between 85% and 90%. The values were derived from various clinical and field specimens when compared with virus isolation with embryonating chicken eggs. On routine AIV surveillance samples, the dot-ELISA achieved a 92%-100% Sp on the basis of testing over 1500 AIV surveillance samples that were confirmed negative by virus isolation. The dot-ELISA detected AIV antigens with a 5-µl allantoic fluid sample that contained a concentration of 0.4 hemagglutinating units. Furthermore, the dot-ELISA retained its specificity for AIV because no cross-reactions were obtained with various other avian viruses. The findings in this study indicated that the dot-ELISA was highly sensitive and specific and comparable with the commercial Directigen® test in the detection of AIV obtained from clinical and field specimens.