Comparison of a Tissue-Culture Virus-Neutralization Test and the Enzyme-Linked Immunosorbent Assay for Measurement of Antibodies to Infectious Bronchitis

Abstract

Two serological tests, the virus-neutralization (VN) test in tissue culture using a tissue-cell-adapted virus and the enzyme-linked immunosorbent assay (ELISA), were compared to detect antibodies against Massachusetts 41 and Connecticut 46 strains of Infectious Bronchitis Virus (IBV). The VN test was conducted in wells of microplates by the usual procedure. The two strains of IBV were adapted after 20 serial passages to induce CPE in 24 hours in chickens embryos kidney cells (CEKC). The ELISA test was carried out using partially virus following ultracentrifugation of each stain of IBV as antigen. The ELISA test detected higher geometric mean antibody titers (GMT) against both strains of IBV than did the VN test. One hundred four serum samples taken at 1, 3, 5, 9, 22, 24, and 26 weeks of age from a flock of chickens vaccinated with the Mass strain three times and the Conn strain of IBV two times during the growing period showed higher antibody titer responses to the Conn 46 than to the Mass 41 strain. Maternal antibodies in chicks one week of age were readily detected by the ELISA test, whereas low or insignificant titers were found by the VN test. Sera of vaccinated chickens collected following challenge with Mass 41 or Conn 46 strain of IBV showed that the ELISA was more sensitive and showed higher titers than did the VN test. Although the VN test showed no rise in GMT in the same sera tested with the heterologous virus, the ELISA showed a slight increase or cross-reaction. The serum samples from the unchallenged control group showed no change in GMT with either test or IBV strain.

Detection of infectious bronchitis virus

“…Reports of indirect ELISAs show them to be group specific (Garcia & Bankowski, 1981;Marquardt et al, 1981;Zellen & Thorsen, 1987;Perrotta et al, 1988;Karaca et al, 1990;De Wit et al, 1997). Antibodies can first be detected by ELISA within 1 week after vaccination or infection (Marquardt et al, 1981;Mockett & Darbyshire, 1981;Hawkes et al, 1983;Monreal et al, 1985;De Wit et al, 1997, 1998a.…”

Coronaviridae: Infectious Bronchitis Virus

“…AGPT is proved to be specific but with poor sensitivity (De Wit et al 1997). ELISA is used on a more frequent basis to measure IBV antibody (Garcia and Bankowski 1981;Marquardt et al 1981;Soula and Moreau 1981;Snyder et al 1985). Among the advantages of ELISA are the increased sensitivity and specificity (Garcia and Bankowski 1981;Marquardt et al 1981;De Wit et al 1997) and the automation of the ELISA steps and calculations (Snyder et al 1983a, b).…”

Detection of Infectious Bronchitis Virus and Specific Anti- Viral Antibodies Using a Concanavalin A–Sandwich–ELISA

“…Enzyme Linked Immunosorbent Assays (ELISAs) have been developed and proved their efficacy, either for the detection of IBV antigen (1,12,15,19,21,34), or anti-IBV antibodies (2,3,4,8,16,22,29).…”

“…Indirect ELISA methods need purified, semi-purified IBV particles, or recombinant IBV antigens for adsorption to the microplate surface (8,16,22,24). Alternatively, polyclonal or monoclonal anti-IBV capture antibodies have been adsorbed on the solid phase in sandwich-ELISA methods for trapping these viruses (12,19,21).…”