Detection of proviral DNA of bovine immunodeficiency virus in bovine tissues by polymerase chain reaction (PCR) and PCR in situ hybridization

Shucheng Zhang, Deryl L. Troyer, Sanjay Kapil, Ling Zheng, George Kennedy, Mark Weiss, Wenzhi Xue, Charles Wood, Harish C. Minocha

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)249-257
Number of pages9
JournalVirology
Volume236
Issue number2
DOIs
StatePublished - Sep 29 1997

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Bovine Immunodeficiency Virus
In Situ Hybridization
Polymerase Chain Reaction
DNA
pol Genes
Tropism
Streptavidin
Lymphoid Tissue
Silver
Gold
Formaldehyde
Esophagus
Smooth Muscle Myocytes
Pancreas
Endothelial Cells
Complementary DNA
Macrophages

ASJC Scopus subject areas

  • Virology

Cite this

Detection of proviral DNA of bovine immunodeficiency virus in bovine tissues by polymerase chain reaction (PCR) and PCR in situ hybridization. / Zhang, Shucheng; Troyer, Deryl L.; Kapil, Sanjay; Zheng, Ling; Kennedy, George; Weiss, Mark; Xue, Wenzhi; Wood, Charles; Minocha, Harish C.

In: Virology, Vol. 236, No. 2, 29.09.1997, p. 249-257.

Research output: Contribution to journalArticle

Zhang, Shucheng ; Troyer, Deryl L. ; Kapil, Sanjay ; Zheng, Ling ; Kennedy, George ; Weiss, Mark ; Xue, Wenzhi ; Wood, Charles ; Minocha, Harish C. / Detection of proviral DNA of bovine immunodeficiency virus in bovine tissues by polymerase chain reaction (PCR) and PCR in situ hybridization. In: Virology. 1997 ; Vol. 236, No. 2. pp. 249-257.
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