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Journal of Clinical Microbiology, April 2005, p. 1843-1845, Vol. 43, No. 4
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.4.1843-1845.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Sequencing and Resolution of Amplified Herpes Simplex Virus DNA with Intermediate Melting Curves as Genotype 1 or 2 by LightCycler PCR Assay

Nicolas C. Issa,1 Mark J. Espy,2 James R. Uhl,2 and Thomas F. Smith2*

Division of Clinical Microbiology, Mayo Clinic College of Medicine, Rochester, Minnesota 55905,2 AIDS Healthcare Foundation, Jacksonville, Florida 322041

Received 3 September 2004/ Returned for modification 25 October 2004/ Accepted 17 December 2004

DNA from 101 specimens containing herpes simplex virus (HSV) produced atypical intermediate melting curves compared with those expected for HSV type 1 or HSV type 2 subsequent to real-time PCR. Nucleic acid sequence analysis of amplified target DNA revealed 1- or 3-bp polymorphisms in the probe region which allowed designation of these viruses as HSV genotype 1 or HSV genotype 2. These two subpopulations of HSV were also identified as HSV genotype 1 or HSV genotype 2 using another commercially available PCR method. Amplified HSV target DNA producing intermediate melting curves could be designated as HSV genotype 1 or HSV genotype 2 without performing sequencing or another PCR method with 96/101 (95%) specimens by adding known intermediate HSV DNA characteristic for the two subpopulations as controls.

* Corresponding author. Mailing address: Division of Clinical Microbiology, Mayo Clinic, 200 First St., SW, Rochester, MN 55905. Phone: (507) 284-8146. Fax: (507) 284-4272. E-mail: tfsmith{at}mayo.edu.

Journal of Clinical Microbiology, April 2005, p. 1843-1845, Vol. 43, No. 4
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.4.1843-1845.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • LeBlanc, J. J., Campbell, S., Pettipas, J., Hatchette, T. F., Davidson, R. J. (2008). Herpes Simplex Virus Type 2 Displays Atypical Melting-Temperature Profiles at Low Viral Titers. J. Clin. Microbiol. 46: 2786-2789 [Abstract] [Full Text]  
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  • Espy, M. J., Uhl, J. R., Sloan, L. M., Buckwalter, S. P., Jones, M. F., Vetter, E. A., Yao, J. D. C., Wengenack, N. L., Rosenblatt, J. E., Cockerill, F. R. III, Smith, T. F. (2006). Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing. Clin. Microbiol. Rev. 19: 165-256 [Abstract] [Full Text]  


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