A PCR-based test for the presence of endogenous virus gene evA in chickens

All endogenous virus, denoted evA, is present at high frequency in all brown egg layer lines. Using inverse polymerase chain reaction (PCR) based on the vital LTR regions, products were obtained containing cellular sequences 5' and 3' to the viral insertion point. PCR of chicken genomic DNA was carried out, using primers chosen from the 5' and 3' cellular sequences and a primer chosen from either the U3 or U5 portions of the viral LTR. Amplification of DNA from birds that did not carry eva with the primer triplets always gave a single 364 hp reaction product, interpreted as representing the flank-to-flank amplification product. Amplification of DNA from known homozygous or heterozygons evA carriers, with the same primer triplets, always gave both the expected junction product and 364 bp product. Therefore, these primer sequences can be used to distinguish eva carriers from non-carriers but cannot distinguish between homozygous and heterozygous evA carriers.