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. 1997 Mar;35(3):548–552. doi: 10.1128/jcm.35.3.548-552.1997

Evaluation of a quantitative competitive PCR assay for measuring herpes simplex virus DNA content in genital tract secretions.

A Hobson 1, A Wald 1, N Wright 1, L Corey 1
PMCID: PMC229624  PMID: 9041386

Abstract

Previous studies have shown an association between the approximate titer of herpes simplex virus (HSV) DNA in clinical specimens and the ability to isolate HSV from genital secretions. To control for variance in amplification conditions, we developed a competitive quantitative PCR (QC PCR) for the detection of HSV DNA. The assay accurately measured from 10 to 10(6) copies of HSV DNA. We compared the QC PCR with our previous semiquantitative detection method and found concordance for 61 of 63 positive specimens. We also evaluated the HSV DNA content from individual swabs of genital secretions obtained from individual sites of the genital tract (cervix, vulva, and rectum) with that from one swab with secretions from all three sites. The concordance for detecting HSV DNA was 91%; for only 4 of 143 collection days was there a > 1 log difference between the two collection methods. A single swab with secretions from all three genital sites and evaluated in a QC PCR format can accurately measure the frequency of subclinical and clinical shedding of HSV and the titer of HSV shed from the genital region. Such an approach should be very useful in the evaluation of antiviral chemotherapy for HSV.

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Selected References

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  1. Becker-André M., Hahlbrock K. Absolute mRNA quantification using the polymerase chain reaction (PCR). A novel approach by a PCR aided transcript titration assay (PATTY). Nucleic Acids Res. 1989 Nov 25;17(22):9437–9446. doi: 10.1093/nar/17.22.9437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Cone R. W., Hobson A. C., Huang M. L. Coamplified positive control detects inhibition of polymerase chain reactions. J Clin Microbiol. 1992 Dec;30(12):3185–3189. doi: 10.1128/jcm.30.12.3185-3189.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Diaz-Mitoma F., Ruben M., Sacks S., MacPherson P., Caissie G. Detection of viral DNA to evaluate outcome of antiviral treatment of patients with recurrent genital herpes. J Clin Microbiol. 1996 Mar;34(3):657–663. doi: 10.1128/jcm.34.3.657-663.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Pachl C., Burke R. L., Stuve L. L., Sanchez-Pescador L., Van Nest G., Masiarz F., Dina D. Expression of cell-associated and secreted forms of herpes simplex virus type 1 glycoprotein gB in mammalian cells. J Virol. 1987 Feb;61(2):315–325. doi: 10.1128/jvi.61.2.315-325.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Peterson E., Schmidt O. W., Goldstein L. C., Nowinski R. C., Corey L. Typing of clinical herpes simplex virus isolates with mouse monoclonal antibodies to herpes simplex virus types 1 and 2: comparison with type-specific rabbit antisera and restriction endonuclease analysis of viral DNA. J Clin Microbiol. 1983 Jan;17(1):92–96. doi: 10.1128/jcm.17.1.92-96.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ramakrishnan R., Fink D. J., Jiang G., Desai P., Glorioso J. C., Levine M. Competitive quantitative PCR analysis of herpes simplex virus type 1 DNA and latency-associated transcript RNA in latently infected cells of the rat brain. J Virol. 1994 Mar;68(3):1864–1873. doi: 10.1128/jvi.68.3.1864-1873.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Wald A., Zeh J., Barnum G., Davis L. G., Corey L. Suppression of subclinical shedding of herpes simplex virus type 2 with acyclovir. Ann Intern Med. 1996 Jan 1;124(1 Pt 1):8–15. doi: 10.7326/0003-4819-124-1_part_1-199601010-00002. [DOI] [PubMed] [Google Scholar]

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