Detection of herpes simplex virus type 1, herpes simplex virus type 2 and varicella-zoster virus in skin lesions. Comparison of real-time PCR, nested PCR and virus isolation

J Clin Virol. 2004 Feb;29(2):120-6. doi: 10.1016/s1386-6532(03)00113-6.

Abstract

Background: Herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2) and varicella-zoster virus (VZV) cause a wide range of signs and symptoms, varying from trivial mucocutaneous lesions to life-threatening infections, especially in immuno-suppressed patients. Since antiviral drugs are available, rapid and sensitive laboratory diagnosis of these virus infections is important.

Objective: To set up and evaluate HSV-1, HSV-2 and VZV qualitative real-time PCR on the Lightcycler system and to compare the results with those of the 'in-house' nested PCR and virus isolation.

Study design: 110 consecutive samples from dermal or genital lesions from patients suspected of having HSV infections and another 110 samples from patients with suspected VZV infections were tested with real-time PCR, nested PCR and virus isolation.

Results: 24 samples (22%) were positive for HSV-1 by virus isolation and nested PCR, whereas 26 (24%) were positive by real-time PCR. HSV-2 was detected in 28 samples (25%) by virus isolation, in 41 (37%) by nested PCR and in 40 (36%) by real-time PCR. VZV was isolated in 15 samples (14%) and VZV DNA was detected in 51 samples (46%) by nested PCR as well as by real-time PCR. Nucleic acid amplification increased the detection rate of HSV-2 and VZV DNA in particular compared to virus isolation. No significant difference in sensitivity was found between real-time PCR and nested PCR.

Conclusion: Real-time PCR has the advantage of rapid amplification, a reduced risk for contamination and it is a suitable method for diagnosis of VZV and HSV in specimens from skin lesions.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chickenpox / diagnosis
  • Chickenpox / virology
  • Clinical Laboratory Techniques / methods
  • DNA, Viral / analysis
  • DNA, Viral / chemistry
  • DNA, Viral / isolation & purification
  • Genes, Viral
  • Herpes Genitalis / diagnosis
  • Herpes Genitalis / virology
  • Herpes Simplex / diagnosis
  • Herpes Simplex / virology
  • Herpesvirus 1, Human / classification
  • Herpesvirus 1, Human / genetics
  • Herpesvirus 1, Human / growth & development
  • Herpesvirus 1, Human / isolation & purification*
  • Herpesvirus 2, Human / classification
  • Herpesvirus 2, Human / genetics
  • Herpesvirus 2, Human / growth & development
  • Herpesvirus 2, Human / isolation & purification*
  • Herpesvirus 3, Human / classification
  • Herpesvirus 3, Human / genetics
  • Herpesvirus 3, Human / growth & development
  • Herpesvirus 3, Human / isolation & purification*
  • Humans
  • Polymerase Chain Reaction*
  • Sensitivity and Specificity
  • Skin Diseases, Viral / diagnosis
  • Skin Diseases, Viral / virology*
  • Transition Temperature

Substances

  • DNA, Viral