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

• Published in: Journal of general virology Vol. 72; no. 5; pp. 1039 - 1044
• Main Authors: Fujinaga, Yukako, Shimada, Masamitu, Okazawa, Kazuhide, Fukushima, Michio, Kato, Ikunoshin, Fujinaga, Kei
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.05.1991; Society for General Microbiology
• Subjects: Base Sequence; Biological and medical sciences; carcinoma; cervix; DNA; DNA Probes, HPV; DNA, Viral - chemistry; DNA, Viral - genetics; Female; Fundamental and applied biological sciences. Psychology; Genes, Viral; Humans; Microbiology; Molecular Sequence Data; Papillomaviridae - genetics; Papillomaviridae - isolation & purification; Polymerase Chain Reaction; Sequence Alignment; Techniques used in virology; Uterine Cervical Neoplasms - microbiology; Virology; Human; Carcinoma; Typing; Papovaviridae; Uterine cervix; Method; Genital diseases; Papillomavirus; Virus; Polymerase chain reaction; Human papillomavirus; DNA; Detection; Woman
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/0022-1317-72-5-1039

1 Department of Molecular Biology, Cancer Research Institute and 2 Department of Obstetrics and Gynaecology, Sapporo Medical College, S1, W17, Chuo-ku, Sapporo 060 and 3 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.