J Gen Virol
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J Gen Virol 72 (1991), 1039-1044; DOI 10.1099/0022-1317-72-5-1039
© 1991 Society for General Microbiology

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Simultaneous detection and typing of genital human papillomavirus DNA using the polymerase chain reaction

Yukako Fujinaga1, Masamitu Shimada1,3, Kazuhide Okazawa1,3, Michio Fukushima1,2, Ikunoshin Kato3 and Kei Fujinaga1

1 Department of Molecular Biology, Cancer Research Institute
and2 Department of Obstetrics and Gynaecology, Sapporo Medical College, S1, W17, Chuo-ku, Sapporo 060
and3 Bio Research Laboratories, Takara Shuzo Co. Ltd, Seta 3-4-1, Otsu, Shiga 520-21, Japan

A simple method has been developed for detecting a broad range of genital human papillomavirus (HPV) types using the polymerase chain reaction (PCR). We utilized two consensus sequence primer pairs within the E6 and E7 open reading frames to amplify HPV DNA; malignant HPV DNA (from HPV-16, -18, -31, -33, -52b and -58) was amplified using the pU-1M/pU-2R primer pair whereas benign HPV DNA (from HPV-6 and -11) was amplified using the pU-31B/pU-2R primer pair. Identification of the amplification product was confirmed by restriction enzyme digestion. In this study, a pU-1M/pU-2R-mediated PCR was successfully applied to 39 cervical carcinoma specimens; HPV-16 was detected in 19 cases, HPV-18 in five cases, HPV-31 in two cases, HPV-33 in two cases, HPV-52b in one case, HPV-58 in three cases, and an unknown type(s) was detected in four cases. Overall, the prevalence of HPV was 84.6%. The results indicate that this detection system is useful for the detection of HPVs not only of known types but also of new types.

Received 30 November 1990; accepted 22 January 1991.


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