Abstract
A liquid phase blocking ELISA (LPB-ELISA) was developed for the detection and measurement of antibodies against infectious bronchitis virus (IBV). The purified and nonpurified virus used as antigen, the capture and detector antibodies, and the chicken hyperimmune sera were prepared and standardized for this purpose. A total of 156 sera from vaccinated and 100 from specific pathogen-free chickens with no recorded contact with the virus were tested. The respective serum titers obtained in the serum neutralization test (SNT) were compared with those obtained in the LPB-ELISA. There was a high correlation (r2 = 0.8926) between the two tests. The LPB-ELISA represents a single test suitable for the rapid detection of antibodies against bronchitis virus in chicken sera, with good sensitivity (88%), specificity (100%) and agreement (95.31%).
Highlights
A liquid phase blocking enzyme-linked immunosorbent assay (ELISA) (LPB-ELISA) was developed for the detection and measurement of antibodies against infectious bronchitis virus (IBV)
The respective serum titers obtained in the serum neutralization test (SNT) were compared with those obtained in the LPB-ELISA
We developed and standardized a liquid phase blocking ELISA (LPB-ELISA), and compared its results with those obtained by the SNT for the detection of neutralizing antibodies against IBV
Summary
The Massachusetts strain (Mass 41) of IBV was propagated by infecting 9-11-dayold embryonated specific pathogen-free (SPF) eggs and the allantoic fluid was harvested as recommended (3). The allantoic fluid collected and pooled from IBV-infected SPF embryonated eggs was clarified by centrifugation at 2,000 g for 20 min at 4oC and submitted to 59,000 g for ultracentrifugation. The fractions collected from the gradient which absorbed at 254 nm (viral RNA) and 280 nm (total protein) were pooled and diluted in TNE buffer and the protein concentration was determined by the method of Hartrée (7). The virus infectivity was titrated in SPF embryonated eggs, as recommended (1). The same IBV strain, replicated in embryonated SPF eggs and clarified at low-speed centrifugation, was used as nonpurified antigen in the LPB-ELISA
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