DETECTION OF PROVIRAL DNA OF BOVINE IMMUNODEFICIENCY VIRUS IN BOVINE-TISSUES BY POLYMERASE CHAIN-REACTION (PCR) AND PCR IN-SITU HYBRIDIZATION

In this study, experiments were designed to investigate the distributi on of bovine immunodeficiency virus (BIV) proviral DNA in the tissues and cells of infected calves by solution-phase polymerase chain reacti on (sP-PCR) and PCR in situ hybridization (PCR-ISH). Total DNA samples extracted from tissues of 10 BIV-infected and 5 uninfected carves wer e amplified by SP-PCR with the primers directed to the BIV conserved p ol gene segment. The identity of the SP-PCR product was confirmed by S outhern hybridization with a BIV pol gene cDNA probe. SP-PCR results d emonstrated that BIV proviral DNA was present predominantly in neural tissues and some lymphoid tissues in BIV-infected calves. It also was detected frequently in other tissues including lung, heart, esophagus, and pancreas. Further investigation on cell location of BIV proviral DNA was performed by in situ amplification of DNA on formalin-fixed ti ssue sections. The amplified DNA was subjected to in situ hybridizatio n with an internal biotinylated probe and detected with streptavidin-g old followed by silver enhancement. Specific BIV proviral DNA signals were observed in neurons, microglial cells, lymphocytes, septal macrop hages, smooth muscle cells, and endothelial cells. On the basis of the se results, we conclude that BIV replicates in a variety of bovine tis sues in vivo and has a broad cell tropism. (C) 1997 Academic Press.