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Comparison Between In Situ Hybridization and Real-time PCR Technique as a Means of Detecting the Integrated Form of Human Papillomavirus 16 in Cervical Neoplasia

Fujii, Takuma MD*; Masumoto, Nobuo MD*; Saito, Miyuki CT*; Hirao, Nobumaru MD*; Niimi, Shinichi PhD†; Mukai, Makio MD‡; Ono, Akiko MD*; Hayashi, Shigenori MD*; Kubushiro, Kaneyuki MD*; Sakai, Eiichi PhD†; Tsukazaki, Katsumi MD*; Nozawa, Shiro MD*

Diagnostic Molecular Pathology: June 2005 - Volume 14 - Issue 2 - pp 103-108
Original Article
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Integration of the human papillomavirus (HPV) genome is thought to be one of the causes of cancer progression. However, there is controversy concerning the physical status of HPV 16 in premalignant cervical lesions, and there have been no reports on the concordance between detection of the integrated form of HPV16 by real-time PCR and by in situ hybridization. We investigated specimens of cervical intraepithelial neoplasia (CIN) and invasive carcinomas for the physical status of HPV 16 by real-time PCR and in situ hybridization. The presence of the integrated form was detected by both real-time PCR and in situ hybridization in zero of four cases of CIN1, three of six cases of CIN2, nine of 27 cases of CIN3, and two of six cases of invasive carcinomas. Integrated HPV 16 was present in some premalignant lesions but was not always present in carcinomas. The concordance rate between the two methods for the detection of the presence of the integrated form was 37 of 43 (86%) cases. Real-time PCR and in situ hybridization were found to be complementary and convenient techniques for determining the physical status of the HPV genome. We conclude that a combination of both methods is a more reliable means of assessing the physical status of the HPV genome in cervical neoplasia.

From the *Department of Obstetrics and Gynecology, Keio University School of Medicine, Tokyo, Japan; †Nihon Gene Research Laboratories Inc., Sendai City, Japan; and ‡Department of Pathology, Keio University School of Medicine, Tokyo, Japan.

Supported by a Grand-in-Aid for Scientific Research (C) and (A) Japan Society for the Promoter of Science and Research Grants for Life Science and Medicine, and the Keio University Medical Science Fund.

Reprints: Takuma Fujii, Department of Obstetrics and Gynecology, Keio University, School of Medicine, Shinanomachi 35, Shinjuku-ku, Tokyo 160-8582, Japan (e-mail: fujiit@sc.itc.keio.ac.jp).

© 2005 Lippincott Williams & Wilkins, Inc.