This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Nishiwaki, M.
Right arrow Articles by Yoshiki, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nishiwaki, M.
Right arrow Articles by Yoshiki, T.

Next Article 

Journal of Clinical Microbiology, April 2008, p. 1161-1168, Vol. 46, No. 4
0095-1137/08/$08.00+0     doi:10.1128/JCM.00793-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Genotyping of Human Papillomaviruses by a Novel One-Step Typing Method with Multiplex PCR and Clinical Applications{triangledown}

Morie Nishiwaki,1* Tomohiro Yamamoto,1 Somako Tone,1 Taichi Murai,1 Tatsuya Ohkawara,2 Takakuni Matsunami,1 Motoiki Koizumi,3 Yoshitake Takagi,1 Jun Yamaguchi,1 Nobuo Kondo,1 Jun Nishihira,4 Takeharu Horikawa,5 and Takashi Yoshiki5

GLab Pathology Center Co., Ltd., Sapporo, Japan,1 Department of Gastroenterology and Hematology, Graduate School of Medicine, Hokkaido University, Sapporo, Japan,2 EVE Women's Clinic, Sapporo, Japan,3 Department of Medical Management and Informatics, Hokkaido Information University, Sapporo, Japan,4 GeneticLab Co., Ltd., Sapporo, Japan5

Received 13 April 2007/ Returned for modification 11 June 2007/ Accepted 21 January 2008

We describe here a rapid, high-throughput genotyping procedure that allows the simultaneous detection of 16 high- and low-risk genital human papillomavirus (HPV) types by multiplex PCR in a single reaction tube. Multiplex PCR is based on the amplification of HPV DNA by sets of HPV genotype-specific primers, and the genotypes of HPV are visually identified by the sizes of amplicons after they are separated by capillary electrophoresis. The procedure does not include a hybridization step with HPV-specific probes and is rapid and labor-saving. We detected all 16 HPV genotypes (types 16, 58, 52, 51, 56, 31, 18, 39, 66, 59, 6, 33, 30, 35, 45, and 11) with a high sensitivity and a high degree of reproducibility. By using this newly developed method, we conducted a pilot study to examine the correlation between the prevalence and genotype distributions of HPV and the cytological group classifications for 547 cervical samples. Compared with the group of samples considered normal (14.7%), there was a significant increase in the prevalence of HPV in women with atypical squamous cells of unknown significance (61.3%), low-grade intraepithelial lesions (75.8%), and high-grade intraepithelial lesions (HSILs) (82.2%). The prevalence and distribution of type 58 were correlated with cytological malignancies, with the highest prevalence in women with HSILs. In conclusion, the novel multiplex PCR method described appears to be highly suitable not only for the screening of cervical cancer precursor lesions but also for the characterization of genotype distributions in large-scale epidemiological studies and HPV vaccination trials.

* Corresponding author. Mailing address: Department of Advanced Pathology Services, GLab Pathology Center Co., Ltd., 28-196, N9-W15, Chuo-ku, Sapporo 0600009, Japan. Phone: 81-116447342. Fax: 81-116447611. E-mail: morie_nishiwaki{at}gene-lab.com

{triangledown} Published ahead of print on 30 January 2008.

Journal of Clinical Microbiology, April 2008, p. 1161-1168, Vol. 46, No. 4
0095-1137/08/$08.00+0     doi:10.1128/JCM.00793-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»