IN-SITU POLYMERASE-CHAIN-REACTION AMPLIFICATION OF HIV-1 DNA IN BRAIN-TISSUE

A direct in situ polymerase chain reaction (IS-PCR) assay is described for the detection of HIV-1 proviral DNA in formalin fixed paraffin em bedded brain tissue. Biotin-16-dUTP is incorporated during the PCR pro cess 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, th e use of an avidin-biotin-alkaline phosphatase complex (ABC) and the a pplication of tyramide signal amplification (TSA) which allows both ch romogenic and fluorescence detection. TSA detection enhances the sensi tivity of IS-PCR, permitting fewer PCR cycles and preserving tissue mo rphology. (C) 1998 Elsevier Science B.V. All rights reserved.