Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Jul;34(7):1654–1659. doi: 10.1128/jcm.34.7.1654-1659.1996

Detection of a transforming fragment of herpes simplex virus type 2 in clinical specimens by PCR. The Canadian Women's HIV Study Group.

G H Guibinga 1, F Coutlée 1, A Kessous 1, C Hankins 1, N Lapointe 1, G Richer 1, J Tousignant 1
PMCID: PMC229089  PMID: 8784564

Abstract

A PCR assay for the sensitive detection of a transforming fragment of herpes simplex virus type 2 (HSV-2) was developed. Oligonucleotide primers were selected in Xho-2, a transforming region of the BglII N fragment of HSV-2. The assay reached a sensitivity endpoint of 10 copies of the Xho-2 subfragment and did not show cross-reactivity with other herpesviruses, including HSV-1. All 42 HSV-2 isolates scored positive in the assay. The Xho-2 PCR assay was evaluated with 216 clinical specimens and the results were compared with those of cell culture. The best protocol for processing specimens contained in viral transport medium included a centrifugation step followed by cell lysis. Of the 107 specimens positive for HSV-2 by culture, 105 were PCR positive (sensitivity, 98.1%). For one of the two falsely negative samples, beta-globin as well as sequences from the HSV-2 DNA polymerase gene could not be amplified. The other sample scored positive in both of these reactions but was indeterminate in duplicate tests by Xho-2 PCR. Two of 109 HSV-2 culture-negative specimens tested positive in the PCR assay (specificity, 98.2%). The latter two samples tested positive in a PCR test for the HSV-2 DNA polymerase gene. This novel tool was shown to be sensitive and specific for HSV-2 sequences and should allow for the investigation of the role of HSV-2 in genital cancers.

Full Text

The Full Text of this article is available as a PDF (373.1 KB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Bauer H. M., Ting Y., Greer C. E., Chambers J. C., Tashiro C. J., Chimera J., Reingold A., Manos M. M. Genital human papillomavirus infection in female university students as determined by a PCR-based method. JAMA. 1991 Jan 23;265(4):472–477. [PubMed] [Google Scholar]
  3. Cameron I. R., Park M., Dutia B. M., Orr A., Macnab J. C. Herpes simplex virus sequences involved in the initiation of oncogenic morphological transformation of rat cells are not required for maintenance of the transformed state. J Gen Virol. 1985 Mar;66(Pt 3):517–527. doi: 10.1099/0022-1317-66-3-517. [DOI] [PubMed] [Google Scholar]
  4. Clark R. B., Pakiz C. B., Hostetter M. K. Synergistic activity of aminoglycoside-beta-lactam combinations against Pseudomonas aeruginosa with an unusual aminoglycoside antibiogram. Med Microbiol Immunol. 1990;179(2):77–86. doi: 10.1007/BF00198528. [DOI] [PubMed] [Google Scholar]
  5. Cone R. W., Hobson A. C., Brown Z., Ashley R., Berry S., Winter C., Corey L. Frequent detection of genital herpes simplex virus DNA by polymerase chain reaction among pregnant women. JAMA. 1994 Sep 14;272(10):792–796. [PubMed] [Google Scholar]
  6. Cone R. W., Hobson A. C., Palmer J., Remington M., Corey L. Extended duration of herpes simplex virus DNA in genital lesions detected by the polymerase chain reaction. J Infect Dis. 1991 Oct;164(4):757–760. doi: 10.1093/infdis/164.4.757. [DOI] [PubMed] [Google Scholar]
  7. Corey L. Laboratory diagnosis of herpes simplex virus infections. Principles guiding the development of rapid diagnostic tests. Diagn Microbiol Infect Dis. 1986 Mar;4(3 Suppl):111S–119S. doi: 10.1016/s0732-8893(86)80049-9. [DOI] [PubMed] [Google Scholar]
  8. Coutlée F., Saint-Antoine P., Olivier C., Vessous-Elbaz A., Voyer H., Berrada F., Bégin P., Giroux L., Viscidi R. Discordance between primer pairs in the polymerase chain reaction for detection of human immunodeficiency virus type 1: a role for taq polymerase inhibitors. J Infect Dis. 1991 Oct;164(4):817–818. doi: 10.1093/infdis/164.4.817. [DOI] [PubMed] [Google Scholar]
  9. Dascal A., Chan-Thim J., Morahan M., Portnoy J., Mendelson J. Diagnosis of herpes simplex virus infection in a clinical setting by a direct antigen detection enzyme immunoassay kit. J Clin Microbiol. 1989 Apr;27(4):700–704. doi: 10.1128/jcm.27.4.700-704.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DiPaolo J. A., Woodworth C. D., Popescu N. C., Koval D. L., Lopez J. V., Doniger J. HSV-2-induced tumorigenicity in HPV16-immortalized human genital keratinocytes. Virology. 1990 Aug;177(2):777–779. doi: 10.1016/0042-6822(90)90548-6. [DOI] [PubMed] [Google Scholar]
  11. DiPaolo J. A., Woodworth C. D., Popescu N. C., Notario V., Doniger J. Induction of human cervical squamous cell carcinoma by sequential transfection with human papillomavirus 16 DNA and viral Harvey ras. Oncogene. 1989 Apr;4(4):395–399. [PubMed] [Google Scholar]
  12. Franco E. L. Cancer causes revisited: human papillomavirus and cervical neoplasia. J Natl Cancer Inst. 1995 Jun 7;87(11):779–780. doi: 10.1093/jnci/87.11.779. [DOI] [PubMed] [Google Scholar]
  13. Galloway D. A., Goldstein L. C., Lewis J. B. Identification of proteins encoded by a fragment of herpes simplex virus type 2 DNA that has transforming activity. J Virol. 1982 May;42(2):530–537. doi: 10.1128/jvi.42.2.530-537.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Galloway D. A., McDougall J. K. The oncogenic potential of herpes simplex viruses: evidence for a 'hit-and-run' mechanism. Nature. 1983 Mar 3;302(5903):21–24. doi: 10.1038/302021a0. [DOI] [PubMed] [Google Scholar]
  15. Galloway D. A., McDougall J. K. Transformation of rodent cells by a cloned DNA fragment of herpes simplex virus type 2. J Virol. 1981 May;38(2):749–760. doi: 10.1128/jvi.38.2.749-760.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Galloway D. A., Nelson J. A., McDougall J. K. Small fragments of herpesvirus DNA with transforming activity contain insertion sequence-like structures. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4736–4740. doi: 10.1073/pnas.81.15.4736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hardy D. A., Arvin A. M., Yasukawa L. L., Bronzan R. N., Lewinsohn D. M., Hensleigh P. A., Prober C. G. Use of polymerase chain reaction for successful identification of asymptomatic genital infection with herpes simplex virus in pregnant women at delivery. J Infect Dis. 1990 Nov;162(5):1031–1035. doi: 10.1093/infdis/162.5.1031. [DOI] [PubMed] [Google Scholar]
  18. Herrington C. S. Human papillomaviruses and cervical neoplasia. II. Interaction of HPV with other factors. J Clin Pathol. 1995 Jan;48(1):1–6. doi: 10.1136/jcp.48.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jenkins F. J., Howett M. K. Characterization of mRNAs that map in the BglII N fragment of the herpes simplex virus type 2 genome. J Virol. 1984 Oct;52(1):99–107. doi: 10.1128/jvi.52.1.99-107.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jones C. Cervical cancer: is herpes simplex virus type II a cofactor? Clin Microbiol Rev. 1995 Oct;8(4):549–556. doi: 10.1128/cmr.8.4.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kessous-Elbaz A., Pelletier M., Cohen E. A., Langelier Y. Retention and expression of the left end subfragment of the herpes simplex virus type 2 BglII N DNA fragment do not correlate with tumorigenic conversion of NIH 3T3 cells. J Gen Virol. 1989 Aug;70(Pt 8):2171–2177. doi: 10.1099/0022-1317-70-8-2171. [DOI] [PubMed] [Google Scholar]
  22. Kimura H., Futamura M., Kito H., Ando T., Goto M., Kuzushima K., Shibata M., Morishima T. Detection of viral DNA in neonatal herpes simplex virus infections: frequent and prolonged presence in serum and cerebrospinal fluid. J Infect Dis. 1991 Aug;164(2):289–293. doi: 10.1093/infdis/164.2.289. [DOI] [PubMed] [Google Scholar]
  23. Langenberg A., Zbanyszek R., Dragavon J., Ashley R., Corey L. Comparison of diploid fibroblast and rabbit kidney tissue cultures and a diploid fibroblast microtiter plate system for the isolation of herpes simplex virus. J Clin Microbiol. 1988 Sep;26(9):1772–1774. doi: 10.1128/jcm.26.9.1772-1774.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lee H. H., Chernesky M. A., Schachter J., Burczak J. D., Andrews W. W., Muldoon S., Leckie G., Stamm W. E. Diagnosis of Chlamydia trachomatis genitourinary infection in women by ligase chain reaction assay of urine. Lancet. 1995 Jan 28;345(8944):213–216. doi: 10.1016/s0140-6736(95)90221-x. [DOI] [PubMed] [Google Scholar]
  25. Lulitanond V., Chantratita W., Thammaprasert K., Nimmanahaeminda K., Matangkasombut P., Yoosook C. Detection of herpes simplex virus type 2 Bgl II N fragment in paraffin-embedded cervical tissue sections using nested polymerase chain reaction. Mol Cell Probes. 1994 Dec;8(6):441–447. doi: 10.1006/mcpr.1994.1063. [DOI] [PubMed] [Google Scholar]
  26. Matlashewski G., Schneider J., Banks L., Jones N., Murray A., Crawford L. Human papillomavirus type 16 DNA cooperates with activated ras in transforming primary cells. EMBO J. 1987 Jun;6(6):1741–1746. doi: 10.1002/j.1460-2075.1987.tb02426.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Matsumoto T., Yamada O., Itagaki A., Ishida S., Kamahora T., Kurimura T. Rapid DNA diagnosis of herpes simplex virus serotypes. J Virol Methods. 1992 Oct;40(1):119–125. doi: 10.1016/0166-0934(92)90013-4. [DOI] [PubMed] [Google Scholar]
  28. Nahass G. T., Goldstein B. A., Zhu W. Y., Serfling U., Penneys N. S., Leonardi C. L. Comparison of Tzanck smear, viral culture, and DNA diagnostic methods in detection of herpes simplex and varicella-zoster infection. JAMA. 1992 Nov 11;268(18):2541–2544. [PubMed] [Google Scholar]
  29. Pilon L., Kessous-Elbaz A., Langelier Y., Royal A. Transformation of NIH 3T3 cells by herpes simplex type 2 BglII n fragment and sub-fragments is independent from induction of mutation at the HPRT locus. Biochem Biophys Res Commun. 1989 Mar 31;159(3):1249–1255. doi: 10.1016/0006-291x(89)92244-4. [DOI] [PubMed] [Google Scholar]
  30. Rogers B. B., Josephson S. L., Mak S. K., Sweeney P. J. Polymerase chain reaction amplification of herpes simplex virus DNA from clinical samples. Obstet Gynecol. 1992 Mar;79(3):464–469. doi: 10.1097/00006250-199203000-00028. [DOI] [PubMed] [Google Scholar]
  31. Saavedra C., Kessous-Elbaz A. Retention of herpes simplex virus type II sequences in bglII n-transformed cells after co-transfection with a selectable marker. EMBO J. 1985 Dec 16;4(13A):3419–3426. doi: 10.1002/j.1460-2075.1985.tb04099.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shimizu C., Shimizu H., Mitsuda T., Tsukuda M., Ichikawa S., Yokota S. One-step determination of herpes simplex virus types I and II by polymerase chain reaction. Mol Cell Probes. 1994 Jun;8(3):193–198. doi: 10.1006/mcpr.1994.1026. [DOI] [PubMed] [Google Scholar]
  33. Shoji H., Koga M., Kusuhara T., Kaji M., Ayabe M., Hino H., Hondo R. Differentiation of herpes simplex virus 1 and 2 in cerebrospinal fluid of patients with HSV encephalitis and meningitis by stringent hybridization of PCR-amplified DNAs. J Neurol. 1994 Aug;241(9):526–530. doi: 10.1007/BF00873514. [DOI] [PubMed] [Google Scholar]
  34. Vonka V., Kanka J., Roth Z. Herpes simplex type 2 virus and cervical neoplasia. Adv Cancer Res. 1987;48:149–191. doi: 10.1016/s0065-230x(08)60692-2. [DOI] [PubMed] [Google Scholar]
  35. Yamakawa Y., Forslund O., Chua K. L., Dillner L., Boon M. E., Hansson B. G. Detection of the BC 24 transforming fragment of the herpes simplex virus type 2 (HSV-2) DNA in cervical carcinoma tissue by polymerase chain reaction (PCR). APMIS. 1994 Jun;102(6):401–406. doi: 10.1111/j.1699-0463.1994.tb04890.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES