Molecular Diagnosis

, Volume 5, Issue 4, pp 279–284 | Cite as

Detection of Herpes Simplex Virus and Varicella- zoster Virus in Clinical Swabs: Frequent Inhibition of PCR as Determined by Internal Controls

  • Guntram Bezold
  • Matthias Volkenandt
  • Petra GottlÖber
  • Ralf-Uwe Peter
Original research/reviews


Background: PCR-based detection of microorganisms is widely used for diagnostic purposes. Most routine PCR applications do not control for inhibition of PCR, thus leading to false-negative results.

Methods and Results: One hundred eighteen swab samples obtained from skin and mucosa were investigated for the presence of herpes simplex virus (HSV), varicellazoster virus (VZV), and the control gene β-globin by internally controlled PCR with purified and unpurified DNA in parallel. With unpurified DNA, inhibition of PCR was detected in 23% of β-globin PCRs, 25% of VZV PCRs, and 16% of HSV PCRs versus 3% each for purified DNA. Approximately 20% of the samples with positive results for HSV or VZV had negative or inhibited results using unpurified DNA.

Conclusion: These results indicate that PCR from clinical swab specimens should be performed exclusively with internal controls because the positive control alone cannot exclude PCR inhibition in individual samples. Purification of DNA will decrease, but not exclude, PCR inhibition.


quality control competitive PCR mimics 


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  1. 1.
    Verkooyen RP, Luijendijk A, Huisman WM, et al.: Detection of PCR inhibitors in cervical specimens by using the Amplicor Chlamydia trachomatis assay. J Clin Microbiol 1996;34:3072–3074PubMedGoogle Scholar
  2. 2.
    Rübben A, Baron JM, Grussendorf-Conen EI: Routine detection of herpes simplex virus and varicella zoster virus by polymerase chain reaction reveals that initial herpes zoster is frequently misdiagnosed as herpes simplex. Br J Dermatol 1997;137:259–261PubMedCrossRefGoogle Scholar
  3. 3.
    Nahass GT, Mandel MJ, Cook S, Fan W, Leonardi CL: Detection of herpes simplex infection from cutaneous lesions in different clinical stages with the polymerase chain reaction. J Am Acad Dermatol 1995;32:730–733PubMedCrossRefGoogle Scholar
  4. 4.
    Cone RW, Hobson AC, 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;164:757–760PubMedCrossRefGoogle Scholar
  5. 5.
    LaRussa P, Steinberg S, Meurice F, Gershon A: Transmission of vaccine strain varicella-zoster virus from a healthy adult with vaccine-associated rash to susceptible household contacts. J Infect Dis 1997;176:1072–1075PubMedCrossRefGoogle Scholar
  6. 6.
    Jackson R, Morris DJ, Cooper RJ, et al.: Multiplex polymerase chain reaction for adenovirus and herpes simplex virus in eye swabs. J Virol Methods 1996;56:41–48PubMedCrossRefGoogle Scholar
  7. 7.
    Furuta Y, Fukuda S, Suzuki S, Takasu T, Inuyama Y, Nagashima K: Detection of varicella-zoster virus DNA in patients with acute peripheral facial palsy by the polymerase chain reaction, and its use for early diagnosis of zoster sine herpete. J Med Virol 1997;52:316–319PubMedCrossRefGoogle Scholar
  8. 8.
    Lichtensteiger CA, Steenbergen SM, Lee RM, Polson DD, Vimr ER: Direct PCR analysis for toxigenic Pasteurella multocida. J Clin Microbiol 1996;34:3035–3039PubMedGoogle Scholar
  9. 9.
    Saiki RK, Gelfand DH, Stoffel S, et al.: Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 1988;239:487–491PubMedCrossRefGoogle Scholar
  10. 10.
    Cao M, Xiao X, Egbert B, Darragh TM, Yen TSB: Rapid detection of cutaneous herpes simplex virus infection with the polymerase chain reaction. J Invest Dermatol 1989;82:391–392CrossRefGoogle Scholar
  11. 11.
    Tsurumi T, Maeno K, Nishigama Y: Nucleotide sequence of the DNA polymerase gene of herpes simplex virus type 2 and comparison with the type 1 counterpart. Gene 1987;52:129–137PubMedCrossRefGoogle Scholar
  12. 12.
    Puchhammer-Stöckl E, Popow-Kraupp T, Heinz FX, Mandl CW, Kunz C: Detection of varicella-zoster virus DNA by polymerase chain reaction in the cerebrospinal fluid of patients suffering from neurological complications associated with chicken pox or herpes zoster. J Clin Microbiol 1991;29:1513–1516PubMedGoogle Scholar
  13. 13.
    Saiki RK, Scharf S, Faloona F, et al.: Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 1985;230:1350–1354PubMedCrossRefGoogle Scholar
  14. 14.
    Siebert PD, Larrick JW: Competitive PCR. Nature 1992;359:557–558PubMedCrossRefGoogle Scholar
  15. 15.
    Cone RW, Hobson AC, Huang MLW: Coamplified positive control detects inhibition of polymerase chain reaction. J Clin Microbiol 1992;30:3185–3189PubMedGoogle Scholar
  16. 16.
    Zerbini M, Gallinella G, Manaresi E, Musiani M, Gentilomi G, Venturoli S: Standardization of a PCR ELISA in serum samples: Diagnosis of active parvovirus B19 infection. J Med Virol 1999;59:239–244PubMedCrossRefGoogle Scholar
  17. 17.
    Thoreson AC, Borre M, Andersen LP, et al.: Helicobacter pylori detection in human biopsies: A competitive PCR assay with internal control reveals false results. FEMS Immunol Med Microbiol 1999;24:201–208PubMedCrossRefGoogle Scholar
  18. 18.
    Lee PY, Mangan J, Holliman RE, Butcher PD: Quantitation of Toxoplasma gondii DNA in a competitive nested polymerase chain reaction. J Clin Pathol 1999;52:61–64PubMedCrossRefGoogle Scholar
  19. 19.
    Lichtinghagen R, Glaubitz R: A competitive polymerase chain reaction assay for reliable identification of Bordetella pertussis in nasopharyngeal swabs. Eur J Clin Chem Clin Biochem 1995;33:87–93PubMedGoogle Scholar
  20. 20.
    Hohlfeld P, Daffos F, Costa JM, Thulliez P, Forestier F, Vidaud M: Prenatal diagnosis of congenital toxoplasmosis with a polymerase chain reaction test on amniotic fluid. N Engl J Med 1994;331:695–699PubMedCrossRefGoogle Scholar

Copyright information

© Churchill Livingstone® 2000

Authors and Affiliations

  • Guntram Bezold
    • 1
  • Matthias Volkenandt
    • 2
  • Petra GottlÖber
    • 1
  • Ralf-Uwe Peter
    • 1
  1. 1.Department of DermatologyUniversity of UlmUlm and MunichGermany
  2. 2.Department of DermatologyLudwig-Maximilians UniversityUlm and MunichGermany

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