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Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1991 Aug;44(8):676–680. doi: 10.1136/jcp.44.8.676

Detection of Epstein-Barr virus genomes in AIDS related lymphomas: sensitivity and specificity of in situ hybridisation compared with Southern blotting.

S J Hamilton-Dutoit 1, H J Delecluse 1, M Raphael 1, G Lenoir 1, G Pallesen 1
PMCID: PMC496764  PMID: 1653789

Abstract

Eighteen cases of AIDS related, non-Hodgkin's lymphomas were examined for the presence of Epstein-Barr virus (EBV) genomes using in situ hybridisation with a 35S-labelled probe. The results were compared with those obtained independently by Southern blot analysis with a 32P-labelled probe of frozen tissue from the same tumours. Technically satisfactory results were obtained with both methods in 15 lymphomas. EBV DNA was detected in seven of 15 (47%) cases by in situ hybridisation and in eight of 15 (53%) cases by Southern blotting (including all the cases positive by in situ hybridisation). The results of EBV DNA detection by the two techniques were identical in 14 of 15 (93%) cases. In situ hybridisation gave no false positive results. This study shows that the sensitivity and specificity of in situ hybridisation for the detection of EBV genomes in AIDS related lymphomas approaches that of Southern blotting, even when using routinely processed archival, paraffin wax embedded material.

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Selected References

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  1. Bashir R. M., Harris N. L., Hochberg F. H., Singer R. M. Detection of Epstein-Barr virus in CNS lymphomas by in-situ hybridization. Neurology. 1989 Jun;39(6):813–817. doi: 10.1212/wnl.39.6.813. [DOI] [PubMed] [Google Scholar]
  2. Bashir R., Hochberg F., Singer R. H. Detection of Epstein-Barr virus by in situ hybridization. Progress toward development of a nonisotopic diagnostic test. Am J Pathol. 1989 Dec;135(6):1035–1044. [PMC free article] [PubMed] [Google Scholar]
  3. Brigati D. J., Myerson D., Leary J. J., Spalholz B., Travis S. Z., Fong C. K., Hsiung G. D., Ward D. C. Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization probes. Virology. 1983 Apr 15;126(1):32–50. doi: 10.1016/0042-6822(83)90460-9. [DOI] [PubMed] [Google Scholar]
  4. EPSTEIN M. A., ACHONG B. G., BARR Y. M. VIRUS PARTICLES IN CULTURED LYMPHOBLASTS FROM BURKITT'S LYMPHOMA. Lancet. 1964 Mar 28;1(7335):702–703. doi: 10.1016/s0140-6736(64)91524-7. [DOI] [PubMed] [Google Scholar]
  5. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  6. Greenspan J. S., Greenspan D., Lennette E. T., Abrams D. I., Conant M. A., Petersen V., Freese U. K. Replication of Epstein-Barr virus within the epithelial cells of oral "hairy" leukoplakia, an AIDS-associated lesion. N Engl J Med. 1985 Dec 19;313(25):1564–1571. doi: 10.1056/NEJM198512193132502. [DOI] [PubMed] [Google Scholar]
  7. Hamilton-Dutoit S. J., Pallesen G., Franzmann M. B., Karkov J., Black F., Skinhøj P., Pedersen C. AIDS-related lymphoma. Histopathology, immunophenotype, and association with Epstein-Barr virus as demonstrated by in situ nucleic acid hybridization. Am J Pathol. 1991 Jan;138(1):149–163. [PMC free article] [PubMed] [Google Scholar]
  8. Hamilton-Dutoit S. J., Pallesen G., Karkov J., Skinhøj P., Franzmann M. B., Pedersen C. Identification of EBV-DNA in tumour cells of AIDS-related lymphomas by in-situ hybridisation. Lancet. 1989 Mar 11;1(8637):554–552. doi: 10.1016/s0140-6736(89)90093-7. [DOI] [PubMed] [Google Scholar]
  9. Harabuchi Y., Yamanaka N., Kataura A., Imai S., Kinoshita T., Mizuno F., Osato T. Epstein-Barr virus in nasal T-cell lymphomas in patients with lethal midline granuloma. Lancet. 1990 Jan 20;335(8682):128–130. doi: 10.1016/0140-6736(90)90002-m. [DOI] [PubMed] [Google Scholar]
  10. Henle G., Henle W., Diehl V. Relation of Burkitt's tumor-associated herpes-ytpe virus to infectious mononucleosis. Proc Natl Acad Sci U S A. 1968 Jan;59(1):94–101. doi: 10.1073/pnas.59.1.94. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ho M., Miller G., Atchison R. W., Breinig M. K., Dummer J. S., Andiman W., Starzl T. E., Eastman R., Griffith B. P., Hardesty R. L. Epstein-Barr virus infections and DNA hybridization studies in posttransplantation lymphoma and lymphoproliferative lesions: the role of primary infection. J Infect Dis. 1985 Nov;152(5):876–886. doi: 10.1093/infdis/152.5.876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lawrence J. B., Villnave C. A., Singer R. H. Sensitive, high-resolution chromatin and chromosome mapping in situ: presence and orientation of two closely integrated copies of EBV in a lymphoma line. Cell. 1988 Jan 15;52(1):51–61. doi: 10.1016/0092-8674(88)90530-2. [DOI] [PubMed] [Google Scholar]
  13. Moar M. H., Klein G. Detection of Epstein-Barr virus (EBV) DNA sequences using in situ hybridization. Biochim Biophys Acta. 1978 Jun 22;519(1):49–64. doi: 10.1016/0005-2787(78)90061-8. [DOI] [PubMed] [Google Scholar]
  14. Murphy J. K., Young L. S., Bevan I. S., Lewis F. A., Dockey D., Ironside J. W., O'Brien C. J., Wells M. Demonstration of Epstein-Barr virus in primary brain lymphoma by in situ DNA hybridisation in paraffin wax embedded tissue. J Clin Pathol. 1990 Mar;43(3):220–223. doi: 10.1136/jcp.43.3.220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Niedobitek G., Hamilton-Dutoit S., Herbst H., Finn T., Vetner M., Pallesen G., Stein H. Identification of Epstein-Barr virus-infected cells in tonsils of acute infectious mononucleosis by in situ hybridization. Hum Pathol. 1989 Aug;20(8):796–799. doi: 10.1016/0046-8177(89)90075-0. [DOI] [PubMed] [Google Scholar]
  16. Nuovo G. J., Richart R. M. Buffered formalin is the superior fixative for the detection of HPV DNA by in situ hybridization analysis. Am J Pathol. 1989 Apr;134(4):837–842. [PMC free article] [PubMed] [Google Scholar]
  17. Nuovo G. J., Silverstein S. J. Comparison of formalin, buffered formalin, and Bouin's fixation on the detection of human papillomavirus deoxyribonucleic acid from genital lesions. Lab Invest. 1988 Nov;59(5):720–724. [PubMed] [Google Scholar]
  18. Pringle J. H., Homer C. E., Warford A., Kendall C. H., Lauder I. In situ hybridization: alkaline phosphatase visualization of biotinylated probes in cryostat and paraffin sections. Histochem J. 1987 Sep;19(9):488–496. doi: 10.1007/BF01675419. [DOI] [PubMed] [Google Scholar]
  19. Rentrop M., Knapp B., Winter H., Schweizer J. Aminoalkylsilane-treated glass slides as support for in situ hybridization of keratin cDNAs to frozen tissue sections under varying fixation and pretreatment conditions. Histochem J. 1986 May;18(5):271–276. doi: 10.1007/BF01676237. [DOI] [PubMed] [Google Scholar]
  20. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  21. Saemundsen A. K., Purtilo D. T., Sakamoto K., Sullivan J. L., Synnerholm A. C., Hanto D., Simmons R., Anvret M., Collins R., Klein G. Documentation of Epstein-Barr virus infection in immunodeficient patients with life-threatening lymphoproliferative diseases by Epstein-Barr virus complementary RNA/DNA and viral DNA/DNA hybridization. Cancer Res. 1981 Nov;41(11 Pt 1):4237–4242. [PubMed] [Google Scholar]
  22. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  23. Stansfeld A. G., Diebold J., Noel H., Kapanci Y., Rilke F., Kelényi G., Sundstrom C., Lennert K., van Unnik J. A., Mioduszewska O. Updated Kiel classification for lymphomas. Lancet. 1988 Feb 6;1(8580):292–293. doi: 10.1016/s0140-6736(88)90367-4. [DOI] [PubMed] [Google Scholar]
  24. Teo C. G., Griffin B. E. Visualization of single copies of the Epstein-Barr virus genome by in situ hybridization. Anal Biochem. 1990 Apr;186(1):78–85. doi: 10.1016/0003-2697(90)90576-u. [DOI] [PubMed] [Google Scholar]
  25. Weiss L. M., Movahed L. A. In situ demonstration of Epstein-Barr viral genomes in viral-associated B cell lymphoproliferations. Am J Pathol. 1989 Mar;134(3):651–659. [PMC free article] [PubMed] [Google Scholar]
  26. Weiss L. M., Movahed L. A., Warnke R. A., Sklar J. Detection of Epstein-Barr viral genomes in Reed-Sternberg cells of Hodgkin's disease. N Engl J Med. 1989 Feb 23;320(8):502–506. doi: 10.1056/NEJM198902233200806. [DOI] [PubMed] [Google Scholar]
  27. Weiss L. M., Strickler J. G., Warnke R. A., Purtilo D. T., Sklar J. Epstein-Barr viral DNA in tissues of Hodgkin's disease. Am J Pathol. 1987 Oct;129(1):86–91. [PMC free article] [PubMed] [Google Scholar]
  28. Ziegler J. L., Beckstead J. A., Volberding P. A., Abrams D. I., Levine A. M., Lukes R. J., Gill P. S., Burkes R. L., Meyer P. R., Metroka C. E. Non-Hodgkin's lymphoma in 90 homosexual men. Relation to generalized lymphadenopathy and the acquired immunodeficiency syndrome. N Engl J Med. 1984 Aug 30;311(9):565–570. doi: 10.1056/NEJM198408303110904. [DOI] [PubMed] [Google Scholar]
  29. zur Hausen H., Schulte-Holthausen H., Klein G., Henle W., Henle G., Clifford P., Santesson L. EBV DNA in biopsies of Burkitt tumours and anaplastic carcinomas of the nasopharynx. Nature. 1970 Dec 12;228(5276):1056–1058. doi: 10.1038/2281056a0. [DOI] [PubMed] [Google Scholar]

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