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Comparison of Human Papillomavirus Detection and Genotyping with Four Different Prime Sets by PCR-Sequencing
Authors:CAI Yu Pin  YANG Yi  ZHU Bao Li  LI Yuan  XIA Xiao Yu  ZHANG Rui Fen  #  and XIANG Yang  #  .CAS
Affiliation:Key Laboratory of Pathogenic Microbiology & Immunology,Institute of Microbiology,the Chinese Academy of Sciences,Beijing 100101,China;2.Department of Obstetrics and Gynecology,Peking Union Medical College Hospital,Peking Union Medical College and the Chinese Academy of Medical Sciences,Beijing 100005,China;3.Graduate University of the Chinese Academy of Sciences,Beijing 100101,China
Abstract:ObjectiveTo assess and compare the Human Papillomavirus (HPV) detection efficiency and the potential clinical utility of PCR sequencing-based technology.MethodsFour HPV consensus primer sets (GP5+/6+, MGP, MY09/11, and PGMY09/11) were used in order to amplify a broad spectrum of HPV types for HPV infection in 325 cervical samples and the PCR products were sequenced afterwards for the HPV genotyping.ResultsThe HPV-positive rate was 75.4%, of which 35.5% harbored more than one HPV genotype. A total of 36 different genotypes was found, with HPV 16 (24.1%) being the most prevalent, followed by HPV 58 (13.3%) and HPV 52 (9.6%). There were substantial to almost perfect agreements between different primer sets regarding HPV detection efficiency, with the kappa value varying from 0.751 to 0.925, MGP, and PGMY09/11 were the most effective in detecting multiple infections (P<0.001). With each of the primer sets, a board range of HPV types could be identified, though there were several differences for a few genotypes.ConclusionThe substantial agreement between PCR-sequencing and HC2 for the detection of high-risk HPV (kappa=0.761) indicated that PCR-sequencing is also suitable for routine HPV screening.
Keywords:Human papillomavirus  Cervical cancer  Polymerase chain reaction  DNA sequencing
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