title: Establishment and Application of a Method for High-Risk Human Papillomavirus Genotyping in Cervical Cancer Tissue.
abstract: BACKGROUND
  Persistent high-risk HPV infection is a major cause of cervical cancer and E6/E7 genes and the Li gene in the HPV genome are key targets to detect high-risk HPV. This study aims to explore the relationship between cervical lesions and E6/7 by establishing a polymerase chain reaction (PCR) to detect multiplex genes based on HPV EE7 genes. It is hoped that such methods will provide a more reliable method for clinical screening and the prevention of cervical cancer.
  
  
  METHODS
  Based on alignment, specific primers were designed for HPV E6/E7 genes, the sequences of which came from five5 high-risk papillomaviruses that are common in China. This enabled an E6/E7 gene detection method based on multiplex PCR to be established. E6/E7 and Li gene testing were then performed on 65 cervical cancer tissue samples. The gene copy number of HPV E6/E7 genes and the Li gene were detected from different classifications by real-time fluorescence quantitative PCR.
  
  
  RESULTS
  Out of the 65 cervical cancer tissue samples, 47 (72.31%) showed positive results in E6/E7 multiplex PCR, 21 (32.31%) showed positive results in the Ll gene PCR, and out of the 219 cervical exfoliate cell samples, 56 (25.57%) showed positive results in E6/E7 multiplex PCR, 21 (13.24%) showed positive results in the L1 gene PCR. There were significant differences (p < 0.05) between these two test results. Fluorescent quantitative PCR showed that the ratio of gene copy number of L1 genes and E6/E7 genes was below 1 (p < 0.05) in cervical cancer tissue, in which both the Li and E6/E7 genes coexist.
  
  
  CONCLUSIONS
  The established HPV multiplex PCR assay based on the design of E6/E7 gene is a specific and sensitive method for the detection and genotype of five high-risk HPVs.
authors.forename: Ruanmin
authors.initials: R
authors.lastname: Zou
authors.forename: Wangkai
authors.initials: W
authors.lastname: Xie
authors.forename: Huijing
authors.initials: H
authors.lastname: Wang
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.forename: Lanlan
authors.initials: L
authors.lastname: Xiao
authors.forename: Shuhui
authors.initials: S
authors.lastname: Ji
authors.forename: Xiangmin
authors.initials: X
authors.lastname: Chen
authors.forename: Lifang
authors.initials: L
authors.lastname: Zhang
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Xue
authors.forename: Jun
authors.initials: J
authors.lastname: Chen
mesh_terms: D000328:Adult; D000368:Aged; D002138:Calibration; D004279:DNA, Viral / Q000235:genetics*; D005260:Female; D018628:Gene Dosage; D005838:Genotype; D061809:Human Papillomavirus DNA Tests / Q000379:methods* / Q000592:standards; D006801:Humans; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000592:standards; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics*; D050725:Papillomavirus E7 Proteins / Q000235:genetics*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology; D011237:Predictive Value of Tests; D012015:Reference Standards; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000821:virology
chemical_list: D004279:DNA, Viral; D050725:Papillomavirus E7 Proteins