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Journal of Clinical Microbiology, May 2003, p. 2135-2137, Vol. 41, No. 5
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.5.2135-2137.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Failure To Genotype Herpes Simplex Virus by Real-Time PCR Assay and Melting Curve Analysis Due to Sequence Variation within Probe Binding Sites

Trevor P. Anderson,1* Anja M. Werno,1 Kirsten A. Beynon,1 and David R. Murdoch1,2

Microbiology Unit, Canterbury Health Laboratories,1 Department of Pathology, Christchurch School of Medicine and Health Sciences, University of Otago, Christchurch, New Zealand2

Received 1 November 2002/ Returned for modification 14 December 2002/ Accepted 10 January 2003

Real-time PCR with melting curve analysis of PCR products is a rapid procedure for detecting and genotyping herpes simplex virus (HSV). When testing mucocutaneous samples for HSV by a real-time PCR assay targeting the DNA polymerase gene, we found that some PCR products had atypical melting curves that did not conform to the expected melting temperatures for HSV type 1 or 2. Sequence analysis showed that these strains had base-pair mismatches over the probe binding sites. An alternative assay is required to type such atypical isolates.

* Corresponding author. Mailing address: Canterbury Health Laboratories, Microbiology Unit, P.O. Box 151, Cnr Tuam St. and Hagley Ave., Christchurch, New Zealand. Phone: 64 (03) 3640 346. Fax: 64 (03) 3640 238. E-mail: Trevor.Anderson{at}cdhb.govt.nz.

Journal of Clinical Microbiology, May 2003, p. 2135-2137, Vol. 41, No. 5
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.5.2135-2137.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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