Skip to main content
Journal of Clinical Laboratory Analysis logoLink to Journal of Clinical Laboratory Analysis
. 2000 Sep 27;14(5):214–219. doi: 10.1002/1098-2825(2000)14:5<214::AID-JCLA3>3.0.CO;2-4

Detection and typing of HSV‐1, HSV‐2, and VZV by a multiplex polymerase chain reaction

Panayotis Markoulatos 1,, Amalia Georgopoulou 1, Constantin Kotsovassilis 2, Paraskevi Karabogia‐Karaphillides 3, Niki Spyrou 1
PMCID: PMC6807757  PMID: 11018799

Abstract

The development of a multiplex polymerase chain reaction method for the rapid and accurate detection and typing of HSV‐1, HSV‐2, and VZV from clinical specimens is described. A sensitive multiplex polymerase chain reaction was achieved by optimization of parameters such as the primers, magnesium, and dNTPs concentrations. False‐negative results that sometimes arise due to inhibitors of DNA amplification or failure of DNA extraction procedure used may be avoided by assaying each specimen with α‐tubulin primers. Multiplex PCR amplified viral sequences from all 55 specimens obtained from patients with clinical evidence of HSV or VZV infection indicated 100% sensitivity. From 55 patients who were investigated by multiplex PCR, HSV‐1 was detected in 28, HSV‐2 in 20, and VZV in 7 specimens. The reported results indicate that the present multiplex PCR assay has a potential application in clinical diagnosis when a rapid and accurate detection and typing of involved viruses HSV‐1, HSV‐2, or VZV is needed. J. Clin. Lab. Anal. 14:214–219, 2000. © 2000 Wiley‐Liss, Inc.

Keywords: polymerase chain reaction (PCR), multiplex, herpes simplex virus types 1 and 2, varicella zoster virus

REFERENCES

  • 1. Penneys NS, Goldstein B, Nahass GT, Leonardi CL, Zhu WY. 1991. Herpes simplex virus DNA in occult lesions: demonstration by the polymerase chain reaction. J Am Acad Dermatol 24:689–692. [DOI] [PubMed] [Google Scholar]
  • 2. Casas I, Tenorio A, Ory F, Lozano A, Echevarria JM. 1996. Detection of both herpes simplex and varicella‐zoster viruses in cerebrospinal fluid from patients with encephalitis. J Med Virol 50:82–92. [DOI] [PubMed] [Google Scholar]
  • 3. De Clerq E, Descamp J, Verhelst G, et al. 1980. Comparative efficacy of antiherpes drugs against different strains of herpes simplex virus. J Infect Dis 141:663–574. [DOI] [PubMed] [Google Scholar]
  • 4. Reeves C, Corey L, Adams H, Vontver L, Holmes K. 1981. Risk of recurrence after first episodes of genital herpes: relation to HSV type and antibody response. N Engl J Med 305:315–319. [DOI] [PubMed] [Google Scholar]
  • 5. WHO Meeting . 1991. Diagnosis of human herpesviruses: memorandum from a WHO meeting. Bull WHO 69:277–283. [PMC free article] [PubMed] [Google Scholar]
  • 6. Nahass GT, Goldstein BA, Zhu WY, Serfling U, Penneys NS, Leonardi CL. 1992. Comparison of Tzanck smear, viral culture and DNA diagnostic methods in detection of herpes simplex and varicella‐zoster infection. JAMA 268:2541–2544. [PubMed] [Google Scholar]
  • 7. Lonsdale DM. 1979. A rapid technique for distinguishing herpes simplex virus type 1 from type 2 by restriction enzyme technology. Lancet i: 849–851. [DOI] [PubMed] [Google Scholar]
  • 8. Markoulatos P, Fountoucidou P, Marinakis G, et al. 1997. Clear detection and typing of herpes simplex virus types 1 and 2 by an indirect Elisa assay: comparison with three different combined methods—capture Elisa, restriction enzymes and polymerase chain reaction. J Clin Lab Anal 11:146–153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9. Nahass GT, Mandel MJ, Cook S, Fan W, Leonardi CL. 1995. Detection of herpes simplex and varicella zoster infection from cutaneous lesions in different clinical stages with the polymerase chain reaction. J Am Acad Dermatol 32:730–733. [DOI] [PubMed] [Google Scholar]
  • 10. Rozenberg F, Lebon P. 1991. Amplification and characterization of herpes virus DNA in cerebrospinal fluid from patients with acute encephalitis. J Clin Microbiol 29:2412–2417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11. Matsumoto T, Yamada O, Itagaki A, Ishida S, Kamahora T, Kurimura T. 1992. Rapid DNA diagnosis of herpes simplex virus serotypes. J Virol Methods 40:119–126. [DOI] [PubMed] [Google Scholar]
  • 12. Shimizu C, Shimizu H, Mitsuda T, Tsukuda M, Ichikawa S, Yokota S. 1994. One step determination of herpes simplex virus type I and II by polymerase chain reaction. Molec Cellul Probes 8:193–198. [DOI] [PubMed] [Google Scholar]
  • 13. Lucotte G, Bathelier C, Lespiaux V, Bali C, Champenois T. 1996. Detection and genotyping of herpes simplex virus type 1 and 2 by polymerase chain reaction. Molec Cellul Probes 9:287–290. [DOI] [PubMed] [Google Scholar]
  • 14. Baron JM, Rübben A, Grubendorf‐Conen FI. 1996. Evaluation of a new general primer pair for rapid detection and differentiation of HSV‐1, HSV‐2 and VZV by polymerase chain reaction. J Med Virol 49:279–282. [DOI] [PubMed] [Google Scholar]
  • 15. Hawrami K, Harper D, Breuer J. 1997. Typing of varicella‐zoster virus by amplification of DNA polymorphisms. J Virol Methods 57:169–174. [DOI] [PubMed] [Google Scholar]
  • 16. Vandevelde C, Verstraete M, Van Beers D. 1990. Fast multiplex polymerase chain reaction on boiled clinical samples for rapid viral diagnosis. J Virol Methods 30:215–228. [DOI] [PubMed] [Google Scholar]
  • 17. Repp R, Rhiel S, Heermann KH, et al. 1993. Genotyping by multiplex polymerase chain reaction for detection of endemic hepatitis B virus transmission. J Clin Microbiol 31:1095–1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18. Markoulatos P, Samara V, Siafakas N, Plakokefalos E, Spyrou N, Moncany M. 1999. Development of a quadriplex polymerase chain reaction for human cytomegalovirus. J Clin Lab Anal 13:99–105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. Markoulatos P, Mangana‐Vougiouka O, Koptopoulos G, Nomikou K, Papadopoulos O. 2000. Detection of sheep poxvirus in skin biopsy samples by a multiplex polymerase chain reaction. J Virol Methods 84:161–167. [DOI] [PubMed] [Google Scholar]
  • 20. Whitley RJ. 1990. Herpes simplex viruses In: Fields BN, Knipe DM, editors. Virology, 2nd ed. vol. 2 New York: Raven Press; p 1843–1888. [Google Scholar]
  • 21. Beards G, Graham C, Pillay D. 1998. Investigation of vesicular rashes from HSV and VZV by PCR. J Med Virol 54:155–157. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Laboratory Analysis are provided here courtesy of Wiley

RESOURCES