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Journal of Clinical Microbiology, November 2006, p. 4157-4162, Vol. 44, No. 11
0095-1137/06/$08.00+0     doi:10.1128/JCM.01762-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Simultaneous Amplification and Identification of 25 Human Papillomavirus Types with Templex Technology{triangledown}

Jian Han,1 David C. Swan,2 Sharon J. Smith,3 Shanjuan H. Lum,1 Susan E. Sefers,3 Elizabeth R. Unger,2 and Yi-Wei Tang3,4*

Genaco Biomedical Products, Inc., Huntsville, Alabama 35805,1 National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333,2 Departments of Pathology,3 Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 372324

Received 25 August 2006/ Returned for modification 8 September 2006/ Accepted 12 September 2006

The majority of existing human papillomavirus (HPV) genotyping assays are based on multiplex PCR using consensus or degenerate primers. We developed a Templex HPV assay that simultaneously detects and identifies 25 common HPV genotypes in a single-tube reaction using type-specific primers for the HPV-specific E6 and E7 genes. The analytical sensitivities of the Templex assay for HPV type 16 (HPV-16), -18, and -56 were 20, 100, and 20 copies per reaction mixture, respectively. The Templex assay provides semiquantitative information on each type when multiple HPV types coexist in one reaction. We tested 109 clinical cervical specimens previously evaluated with the Digene HC2 high-risk HPV DNA test and found 95.4% concordance between the assay results. The Templex assay provided type-specific results and found multiple types in 29.2% (14 of 48) of high-risk HPV-positive samples. The entire Templex procedure, including DNA extraction, can be completed within 5 hours, providing a rapid and reliable diagnostic tool for HPV detection and typing that is amenable to automation.

* Corresponding author. Mailing address: Molecular Infectious Disease Laboratory, Vanderbilt University Hospital, 4605 TVC, Nashville, TN 37232-5310. Phone: (615) 322-2035. Fax: (615) 343-8420. E-mail: yiwei.tang{at}vanderbilt.edu.

{triangledown} Published ahead of print on 27 September 2006.

Journal of Clinical Microbiology, November 2006, p. 4157-4162, Vol. 44, No. 11
0095-1137/06/$08.00+0     doi:10.1128/JCM.01762-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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