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 distribution of bovine immunodeficiency virus (BIV) proviral DNA in the tissues and cells of infected calves by solution-phase polymerase chain reaction (SP-PCR) and PCR in situ hybridization (PCR-ISH). Total DNA samples extracted from tissues of 10 BIV-infected and 5 uninfected calves were amplified by SP-PCR with the primers directed to the BIV conserved pol gene segment. The identity of the SP-PCR product was confirmed by Southern hybridization with a BIV pol gene cDNA probe. SP-PCR results demonstrated 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 tissue sections. The amplified DNA was subjected to in situ hybridization with an internal biotinylated probe and detected with streptavidin-gold followed by silver enhancement. Specific BIV proviral DNA signals were observed in neurons, microglial cells, lymphocytes, septal macrophages, smooth muscle cells, and endothelial cells. On the basis of these results, we conclude that BIV replicates in a variety of bovine tissues in vivo and has a broad cell tropism.
Zhang
S
S
Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan 66506, USA.
Troyer
D L
DL
Kapil
S
S
Zheng
L
L
Kennedy
G
G
Weiss
M
M
Xue
W
W
Wood
C
C
Minocha
H C
HC
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification*; D005069:Evaluation Studies as Topic; D015751:Genes, pol; D016087:Immunodeficiency Virus, Bovine / Q000235:genetics* / Q000302:isolation & purification*; D017403:In Situ Hybridization / Q000379:methods* / Q000706:statistics & numerical data; D016180:Lentivirus Infections / Q000821:virology*; D009420:Nervous System / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D011533:Proviruses / Q000235:genetics* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014018:Tissue Distribution
D017931:DNA Primers; D004279:DNA, Viral