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

• Published in: Journal of oral pathology & medicine 2000-03, Vol.29 (3), p.118-122
• Main Authors: Mabruk, M. J. E. M. F., Antonio, M., Flint, S. R., Coleman, D. C., Toner, M., Kay, E., Leader, M., Atkins, G. J.
• Format: Article
• Language: English
• Subjects: AIDS-Related Opportunistic Infections - virology; AIDS/HIV; Biological and medical sciences; Biotechnology; Biotin; Case-Control Studies; Dentistry; DNA Probes; DNA, Viral - analysis; Female; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Herpesvirus 4, Human - genetics; Herpesvirus 4, Human - isolation & purification; HIV Seropositivity - virology; Humans; In Situ Hybridization; In Situ Nick-End Labeling; In vitro gene amplification. Pcr technique; Leukoplakia, Hairy - diagnosis; Leukoplakia, Hairy - etiology; Leukoplakia, Hairy - virology; Male; Medical sciences; Methods. Procedures. Technologies; Non tumoral diseases; oral hairy leukoplakia; Otorhinolaryngology. Stomatology; Random Amplified Polymorphic DNA Technique; Sensitivity and Specificity; Upper respiratory tract, upper alimentary tract, paranasal sinuses, salivary glands: diseases, semeiology
• ISSN: 0904-2512; 1600-0714
• DOI: 10.1034/j.1600-0714.2000.290303.x

: 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. All control specimens were negative for EBV‐DNA.