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Journal of Clinical Microbiology, March 2002, p. 863-867, Vol. 40, No. 3
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.3.863-867.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Rapid and Sensitive Detection of Physical Status of Human Papillomavirus Type 16 DNA by Quantitative Real-Time PCR

Shoji Nagao,1 Mitsuo Yoshinouchi,1* Yasunari Miyagi,1 Atsushi Hongo,1 Junichi Kodama,1 Sachio Itoh,2 and Takafumi Kudo1

Department of Obstetrics and Gynecology,,1 Department of Molecular Genetics, Institute of Cellular and Molecular Biology, Okayama University Medical School, Okayama, Japan2

Received 29 May 2001/ Returned for modification 9 August 2001/ Accepted 6 December 2001

A rapid quantitative real-time PCR method was employed to quantify the copy number of E2 and E6 genes for analysis of the physical status of human papillomavirus type 16 (HPV-16) DNA. Significant differences with respect to both copy numbers were found when more than 40% of HPV-16 DNA was integrated with disruption of the E2 gene in an experimental model. The physical status of HPV-16 DNA in 50 clinical samples was exclusively episomal in 21 cases (42%), concomitant in 14 cases (28%), and integrated in 15 cases (30%). The prevalence of integrated and/or concomitant forms of HPV-16 DNA increased with progression of cervical disease. Four of 11 cervical intraepithelial neoplasia involved integrated forms of HPV-16 DNA partially or exclusively. This rapid, sensitive technique is useful in the analysis of the physical status of HPV DNA.

* Corresponding author. Mailing address: Department of Obstetrics and Gynecology, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan. Phone: (81) 86 235 7320. Fax: (81) 86 225 9570. E-mail: m_yoshi{at}cc.okayama-u.ac.jp.

Journal of Clinical Microbiology, March 2002, p. 863-867, Vol. 40, No. 3
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.3.863-867.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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