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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.
Authors:Takuma Fujii  Nobuo Masumoto  Miyuki Saito  Nobumaru Hirao  Shinichi Niimi  Makio Mukai  Akiko Ono  Shigenori Hayashi  Kaneyuki Kubushiro  Eiichi Sakai  Katsumi Tsukazaki  Shiro Nozawa
Institution:Department of Obstetrics and Gynecology, Keio University School of Medicine, Tokyo 160-8582, Japan. fujiit@sc.itc.keio.ac.jp
Abstract: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.
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