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J Virol Methods. 1998 Feb;70(2):119-27.

In situ polymerase chain reaction amplification of HIV-1 DNA in brain tissue.

Author information

1
Department of Pathology, University of Edinburgh, Western General Hospital, UK.

Abstract

A direct in situ polymerase chain reaction (IS-PCR) assay is described for the detection of HIV-1 proviral DNA in formalin fixed paraffin embedded brain tissue. Biotin-16-dUTP is incorporated during the PCR process and microwave pretreatment of tissue sections ensures that no non-specific incorporation into damaged or nicked genomic DNA occurs. Two methods are compared to detect the biotinylated amplified product, the use of an avidin-biotin-alkaline phosphatase complex (ABC) and the application of tyramide signal amplification (TSA) which allows both chromogenic and fluorescence detection. TSA detection enhances the sensitivity of IS-PCR, permitting fewer PCR cycles and preserving tissue morphology.

PMID:
9562406
DOI:
10.1016/s0166-0934(97)00156-0
[Indexed for MEDLINE]

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