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. 1995 Aug;48(8):733–736. doi: 10.1136/jcp.48.8.733

Retrospective determination of HIV-1 status by a PCR method on paraffin wax embedded sections.

T Slavik 1, M Wolfaardt 1, H van Zyl 1, I W Simson 1
PMCID: PMC502800  PMID: 7560200

Abstract

AIM--To develop a simple but reliable polymerase chain reaction (PCR) method to determine the HIV-1 status of patients on formalin fixed, paraffin wax embedded lymph node tissue. METHODS--Fifty lymph node specimens, 20 from HIV-1 seropositive and 30 from HIV-1 seronegative patients, were analysed. Lymph nodes with a variety of disease conditions were included in the study. Tissue sections were treated with a DNA extraction buffer containing proteinase K and the crude cell lysate was used in PCR analysis. Nested primers were used to amplify HIV-1 DNA sequences coding for gag, pol and env proteins. PCR products were demonstrated by polyacrylamide gel electrophoresis. Results were then compared with HIV-1 serology of the patients from whom the tissue was obtained. RESULTS--The PCR method yielded a specificity of 100%, a sensitivity of 95%, a positive predictive value of 100%, and a negative predictive value of 97% when compared with HIV-1 serology. The kappa statistic (0.958) showed an excellent agreement between the PCR method and serology. Furthermore, HIV-1 DNA was demonstrated in lymph node tissue from a serologically unconfirmed acquired immunodeficiency syndrome case necropsied in 1982. CONCLUSION--This PCR method is a simple and reliable means of retrospectively determining the HIV-1 status of patients using formalin fixed, paraffin wax embedded lymph node tissue.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ahmed T., Wormser G. P., Stahl R. E., Mamtani R., Cimino J., Glasser M., Mittelman A., Friedland M., Arlin Z. Malignant lymphomas in a population at risk for acquired immune deficiency syndrome. Cancer. 1987 Aug 15;60(4):719–723. doi: 10.1002/1097-0142(19870815)60:4<719::aid-cncr2820600402>3.0.co;2-u. [DOI] [PubMed] [Google Scholar]
  2. Albert J., Fenyö E. M. Simple, sensitive, and specific detection of human immunodeficiency virus type 1 in clinical specimens by polymerase chain reaction with nested primers. J Clin Microbiol. 1990 Jul;28(7):1560–1564. doi: 10.1128/jcm.28.7.1560-1564.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bootman J. S., Kitchin P. A. An international collaborative study to assess a set of reference reagents for HIV-1 PCR. J Virol Methods. 1992 Apr;37(1):23–41. doi: 10.1016/0166-0934(92)90018-9. [DOI] [PubMed] [Google Scholar]
  4. Defer C., Agut H., Garbarg-Chenon A., Moncany M., Morinet F., Vignon D., Mariotti M., Lefrère J. J. Multicentre quality control of polymerase chain reaction for detection of HIV DNA. AIDS. 1992 Jul;6(7):659–663. doi: 10.1097/00002030-199207000-00007. [DOI] [PubMed] [Google Scholar]
  5. Fox C. H., Hoover S., Currall V. R., Bahre H. J., Cottler-Fox M. HIV in infected lymph nodes. Nature. 1994 Jul 28;370(6487):256–256. doi: 10.1038/370256a0. [DOI] [PubMed] [Google Scholar]
  6. Giri A. A., Lillo F. B., McDermott J. L., Jannuzzi C., Risso S., Fornia G. L., Concedi D. R., Varnier O. E. Detection of HIV-1 sequences in children using radioactive and colorimetric polymerase chain reactions. J Med Virol. 1994 Apr;42(4):414–419. doi: 10.1002/jmv.1890420415. [DOI] [PubMed] [Google Scholar]
  7. Jackson D. P., Lewis F. A., Taylor G. R., Boylston A. W., Quirke P. Tissue extraction of DNA and RNA and analysis by the polymerase chain reaction. J Clin Pathol. 1990 Jun;43(6):499–504. doi: 10.1136/jcp.43.6.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kashala O., Marlink R., Ilunga M., Diese M., Gormus B., Xu K., Mukeba P., Kasongo K., Essex M. Infection with human immunodeficiency virus type 1 (HIV-1) and human T cell lymphotropic viruses among leprosy patients and contacts: correlation between HIV-1 cross-reactivity and antibodies to lipoarabinomannan. J Infect Dis. 1994 Feb;169(2):296–304. doi: 10.1093/infdis/169.2.296. [DOI] [PubMed] [Google Scholar]
  9. Kemp D. J., Smith D. B., Foote S. J., Samaras N., Peterson M. G. Colorimetric detection of specific DNA segments amplified by polymerase chain reactions. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2423–2427. doi: 10.1073/pnas.86.7.2423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kiene P., Milde-Langosch K., Runkel M., Schulz K., Löning T. A simple and rapid technique to process formalin-fixed, paraffin-embedded tissues for the detection of viruses by the polymerase chain reaction. Virchows Arch A Pathol Anat Histopathol. 1992;420(3):269–273. doi: 10.1007/BF01600280. [DOI] [PubMed] [Google Scholar]
  11. Kiyabu M. T., Shibata D., Arnheim N., Martin W. J., Fitzgibbons P. L. Detection of human papillomavirus in formalin-fixed, invasive squamous carcinomas using the polymerase chain reaction. Am J Surg Pathol. 1989 Mar;13(3):221–224. doi: 10.1097/00000478-198903000-00007. [DOI] [PubMed] [Google Scholar]
  12. Krivine A., Firtion G., Cao L., Francoual C., Henrion R., Lebon P. HIV replication during the first weeks of life. Lancet. 1992 May 16;339(8803):1187–1189. doi: 10.1016/0140-6736(92)91131-q. [DOI] [PubMed] [Google Scholar]
  13. Longo M. C., Berninger M. S., Hartley J. L. Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions. Gene. 1990 Sep 1;93(1):125–128. doi: 10.1016/0378-1119(90)90145-h. [DOI] [PubMed] [Google Scholar]
  14. Päbo S. Ancient DNA: extraction, characterization, molecular cloning, and enzymatic amplification. Proc Natl Acad Sci U S A. 1989 Mar;86(6):1939–1943. doi: 10.1073/pnas.86.6.1939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rogers B. B., Alpert L. C., Hine E. A., Buffone G. J. Analysis of DNA in fresh and fixed tissue by the polymerase chain reaction. Am J Pathol. 1990 Mar;136(3):541–548. [PMC free article] [PubMed] [Google Scholar]
  16. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  17. Selik R. M., Starcher E. T., Curran J. W. Opportunistic diseases reported in AIDS patients: frequencies, associations, and trends. AIDS. 1987 Sep;1(3):175–182. [PubMed] [Google Scholar]
  18. Shibata D. K., Arnheim N., Martin W. J. Detection of human papilloma virus in paraffin-embedded tissue using the polymerase chain reaction. J Exp Med. 1988 Jan 1;167(1):225–230. doi: 10.1084/jem.167.1.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Smith S. A., Easton D. F., Evans D. G., Ponder B. A. Allele losses in the region 17q12-21 in familial breast and ovarian cancer involve the wild-type chromosome. Nat Genet. 1992 Oct;2(2):128–131. doi: 10.1038/ng1092-128. [DOI] [PubMed] [Google Scholar]
  20. Sönnerborg A., Abens J., Johansson B., Strannegård O. Detection of human immunodeficiency virus-1 by polymerase chain reaction and virus cultivation. J Med Virol. 1990 Jul;31(3):234–240. doi: 10.1002/jmv.1890310311. [DOI] [PubMed] [Google Scholar]

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