A simple and rapid technique for the detection of Epstein-Barr virus DNA in HIV-associated oral hairy leukoplakia biopsies

A method of generating nucleic acid probes by polymerase chain reaction (PC R) for the detection of Epstein-Barr virus (EBV)-DNA by in situ hybridizati on in oral hairy leukoplakia (OHL) lesions is described. This method has th e advantage over older methods of being cheaper, quicker and retaining sens itivity and specificity. Purified PCR products of Epstein-Barr virus DNA of 110 bp and 328 bp were labelled with biotin by nick translation or random primer labelling and were compared in in situ hybridization experiments wit h probes prepared by incorporation of biotin-labelled nucleotides in the PC R reaction mixture, with EBV viral DNA as a template. These probes were app lied to 18 OHL tongue biopsies known to be positive for EBV-DNA, using a co mmercially available biotin-labelled BamHI "V" fragment EBV-DNA probe. To d etermine the specificity of the probes, we applied them to 20 normal tongue tissue samples and to 12 biopsies taken from keratotic tongue lesions from patients without risk factors for HIV infection and known to be negative f or EBV-DNA. Clear positive signals for EBV-DNA were detected in all 18 case s of OHL biopsies using the amplimer of 328 bp labelled by PCR and random p rimer labelling. However, nick translation labelling was less efficient and sensitive. All control specimens were negative for EBV-DNA.