Specific delineation of BK polyomavirus in kidney tissue with a digoxigenin-labeled DNA probe.

Details

Serval ID
serval:BIB_C1BCF2EE7F0C
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Specific delineation of BK polyomavirus in kidney tissue with a digoxigenin-labeled DNA probe.
Journal
Molecular and Cellular Probes
Author(s)
Parkin R.K., Boeckh M.J., Erard V., Huang M.L., Myerson D.
ISSN
0890-8508
Publication state
Published
Issued date
2005
Peer-reviewed
Oui
Volume
19
Number
2
Pages
87-92
Language
english
Abstract
The detection of BK polyomavirus (BK virus, BKV) in kidney tissue is hampered by nonspecificity of antibodies suited to immunohistochemistry, and nonspecific background with in situ hybridization. The biotin-labeled DNA probe that is commercially available (Enzo Life Sciences, Inc.) shows good signal, but the intrinsic background in kidney tissue is high. We determined that the intrinsic background is due to endogenous biotin or biotin-binding activity in the renal tubular epithelium. Neither antibody blocking procedures nor an avidin/biotin block were entirely satisfactory for eliminating this background staining. We developed a digoxigenin-labeled DNA probe, and protocol, for detecting BK virus in formalin-fixed, paraffin embedded, kidney tissue obtained at autopsy. The hybridization signal is strong and there is no perceptible background staining. Eleven negative control kidneys all failed to hybridize. Conditions for low stringency hybridization may be employed, detecting both the related JC polyomavirus and BKV. Alternatively, high stringency hybridization conditions may be utilized, detecting BKV only. BK associated tubular necrosis is clearly demonstrated in two cases of BK nephritis.
Keywords
Autopsy, BK Virus/genetics, BK Virus/isolation & purification, DNA Probes/genetics, DNA, Viral/genetics, Digoxigenin/chemistry, Humans, Immunohistochemistry, In Situ Hybridization, Kidney/virology, Polyomavirus Infections/virology
Pubmed
Web of science
Create date
02/02/2010 18:03
Last modification date
20/08/2019 16:36
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