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

M. J.E.M.F. Mabruk, M. Antonio, S. R. Flint, D. C. Coleman, M. Toner, E. Kay, M. Leader, G. J. Atkins

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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 BamHI '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. control specimens were negative for EBV-DNA.

Original languageEnglish
Pages (from-to)118-122
Number of pages5
JournalJournal of Oral Pathology and Medicine
Volume29
Issue number3
DOIs
Publication statusPublished - 2000

Fingerprint

Hairy Leukoplakia
Human Herpesvirus 4
HIV
Biopsy
DNA
Biotin
Polymerase Chain Reaction
Tongue
In Situ Hybridization
Hairy Tongue
Nucleic Acid Probes
DNA Probes
Viral DNA
HIV Infections
Nucleotides
Sensitivity and Specificity

Keywords

  • In situ hybridization
  • Oral hairy leukoplakia

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Oral Surgery
  • Otorhinolaryngology
  • Cancer Research
  • Periodontics

Cite this

A simple and rapid technique for the detection of Epstein-Barr virus DNA in HIV-associated oral hairy leukoplakia biopsies. / Mabruk, M. J.E.M.F.; Antonio, M.; Flint, S. R.; Coleman, D. C.; Toner, M.; Kay, E.; Leader, M.; Atkins, G. J.

In: Journal of Oral Pathology and Medicine, Vol. 29, No. 3, 2000, p. 118-122.

Research output: Contribution to journalArticle

Mabruk, M. J.E.M.F. ; Antonio, M. ; Flint, S. R. ; Coleman, D. C. ; Toner, M. ; Kay, E. ; Leader, M. ; Atkins, G. J. / A simple and rapid technique for the detection of Epstein-Barr virus DNA in HIV-associated oral hairy leukoplakia biopsies. In: Journal of Oral Pathology and Medicine. 2000 ; Vol. 29, No. 3. pp. 118-122.
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