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A simple and rapid technique for the detection of Epstein‐Barr virus DNA in HIV‐associated oral hairy leukoplakia biopsies
Author(s) -
Mabruk M. J. E. M. F.,
Antonio M.,
Flint S. R.,
Coleman D. C.,
Toner M.,
Kay E.,
Leader M.,
Atkins G. J.
Publication year - 2000
Publication title -
journal of oral pathology and medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.887
H-Index - 83
eISSN - 1600-0714
pISSN - 0904-2512
DOI - 10.1034/j.1600-0714.2000.290303.x
Subject(s) - nick translation , primer (cosmetics) , biology , polymerase chain reaction , dna , microbiology and biotechnology , virus , biotin , epstein–barr virus , nucleic acid thermodynamics , in situ hybridization , hybridization probe , virology , chemistry , gene , messenger rna , biochemistry , organic chemistry , base sequence
A method of generating nucleic acid probes by polymerase chain reaction (PCR) for the detection of Epstein–Barr virus (EBV)‐DNA by in situ hybridization in oral hairy leukoplakia (OHL) lesions is described. This method has the advantage over older methods of being cheaper, quicker and retaining sensitivity 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 with probes prepared by incorporation of biotin‐labelled nucleotides in the PCR reaction mixture, with EBV viral DNA as a template. These probes were applied to 18 OHL tongue biopsies known to be positive for EBV‐DNA, using a commercially available biotin‐labelled Bam HI “V” fragment EBV‐DNA probe. To determine 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 for EBV‐DNA. Clear positive signals for EBV‐DNA were detected in all 18 cases of OHL biopsies using the amplimer of 328 bp labelled by PCR and random primer labelling. However, nick translation labelling was less efficient and sensitive. All control specimens were negative for EBV‐DNA.