title: MGB-based one-step multiplex real-time PCR method for rapid detection of HPV.
abstract: OBJECTIVE
  For full-scale analysis of Human Papillomavirus (HPV) status in humans, two minor groove binder (MGB)-based one-step multiplex real-time PCR systems were developed: one to screen 16 high-risk HPV (HR-HPV) types, and one to screen a broader spectrum of HPV types (common HPV or C-HPV).
  
  
  METHODS
  Sensitivity and specificity were evaluated using diluted reference plasmids and 20 control human DNA samples. For clinical evaluation, 510 cervical scrape samples were evaluated.
  
  
  RESULTS
  The sensitivity assays revealed that the C-HPV detection system could detect 10 ~ 100 plasmid copies/reaction, while the HR-HPV detection system detected 10 ~ 500 copies. The specificity test revealed that the systems did not yield positive signals from the 20 human genomic DNA samples. Performance was tested on 510 usable clinical samples. The HR-HPV results were compared to those from the Hybrid Capture 2 (HC2) test, which assesses 13 HR-HPV types; the concordance level between the two methods was 90.8% with a kappa value of 0.813.
  
  
  CONCLUSIONS
  These results showed that our novel MGB-based one-step multiplex real-time PCR method may be used for the diagnosis and mass screening of HPV in clinical and large-scale epidemiological studies.
authors.affiliation: Department of In Vitro Diagnostic Reagents and Medium, National Institute for the Control of Pharmaceutical and Biological Products, Beijing, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Huang
authors.forename: Chun
authors.initials: C
authors.lastname: Gao
authors.forename: Xilai
authors.initials: X
authors.lastname: Ding
authors.forename: Shoufang
authors.initials: S
authors.lastname: Qu
authors.forename: Licheng
authors.initials: L
authors.lastname: Liu
authors.forename: Weili
authors.initials: W
authors.lastname: Wu
authors.forename: Lining
authors.initials: L
authors.lastname: Zhang
authors.forename: Jinyin
authors.initials: J
authors.lastname: Zhao
authors.forename: Shangxian
authors.initials: S
authors.lastname: Gao
mesh_terms: D002584:Cervix Uteri / Q000821:virology; D015344:DNA Probes, HPV / Q000235:genetics; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D061809:Human Papillomavirus DNA Tests / Q000379:methods*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D015344:DNA Probes, HPV; D004279:DNA, Viral