A longitudinal study of a novel dot-enzyme-linked immunosorbent assay for detection of avian influenza virus.
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 resting over 1500 AIV surveillance samples that were confirmed negative by virus isolation. The dot-ELISA detected AIV antigens with a 5-microl 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.
Lu
Huaguang
H
Animal Diagnostic Laboratory, Department of Veterinary Science, The Pennsylvania State University, University Park, PA 16802, USA.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis / Q000276:immunology; D002642:Chick Embryo; D002645:Chickens / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D009980:Influenza A virus / Q000145:classification / Q000276:immunology* / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis / Q000821:virology; D008137:Longitudinal Studies; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral