Adaptation of enzyme-linked immunosorbent assay to the avian system

Abstract

A microplate enzyme-linked immunosorbent assay was developed to detect chicken anti-reovirus antibodies. Studies of the parameters which affect the outcome of the assay with avian serum revealed two aspects for a successful assay. First, enzyme-antibody conjugates prepared by the periodate oxidation technique were found to have retained far more immunological activity than conjugates produced by a glutaraldehyde cross-linking. Second, the results indicated an unusually high affinity of chicken immunoglobulin for the microplate plastic which was mostly eliminated by a pretreatment technique with fixed fetal calf serum. The enzyme-linked immunosorbent assay compared favorably with the latex passive agglutination test, yielding a titration endpoint of 1:51,000, or approximately 1,300 times more sensitive than the latex passive agglutination assay. The assay proved not only to be sensitive to less than 1 ng of specific antibody, but also to have low to moderate variance and high reliability. According to Schuurs and van Weemen (13), the number of publications concerning various

Rapid characterization of avian reoviruses using phylogenetic analysis, reverse transcription-polymerase chain reaction and restriction enzyme fragment length polymorphism

“…Several methods have been used to identify ARV isolates and detect antibodies against ARV, including immunodiffusion, virus neutralization, enzymelinked immunosorbent assay, in situ hybridization, in situ RT-PCR, and immunoblot assays (Sahu & Olson, 1975;Slaght et al, 1978Slaght et al, , 1979Lee et al, 1994;Liu et al, 1999aLiu et al, ,b, 2000Liu et al, , 2002. ARV has multiple serotypes, and new variant serotypes continue to be recognized (Takase et al, 1987;Giambrone & Solano, 1988;Lee et al, 1992).…”

Antigen requirements and specificity of a microplate enzyme-linked immunosorbent assay (ELISA) for detecting infectious bronchitis viral antibodies in chicken serum

“…SLAGItT et al (26) demonstrated recently the high affinity of avian immunoglobulins at neutral pH for polystyrene and a possible link between chicken immunoglobulin concentration and adsorption. Moreover the IgG and IgM concentrations increase up to three weeks after the primary IBV inoculation in chickens (12).…”

Relevance of detection of immunoglobulin M antibody response in birds used for arbovirus surveillance

“…Two reports of ELISAs with sera from experimentally infected chickens were found and contain some puzzling results. One, by Slaght et al (9), indicates that avian antibodies exhibit generally lower affinity constants than do mammalian antibodies, require unusually high salt concentrations or low pH for precipitin assays, and have a high affinity for plastic. In the MAC ELISA and IgG assay reported here no attempts were made to address such problems; we simply adapted standard ELISAs used for the detection of IgM and IgG antibodies to the EEE, WEE, and other viruses in sera from humans and equines.…”