title: Genotyping of 11 human papillomaviruses by multiplex PCR with a GeXP analyzer.
abstract: A new, rapid, and high-throughput method was developed for simultaneous detection of 11 human papillomavirus (HPV) genotypes including nine high-risk types (HPV16, 18, 31, 33, 35, 39, 52, 58, and 66) and two low-risk types (HPV6 and 11) in a single tube by multiplex PCR based on a GenomeLab Gene Expression Profiler (GeXP) analyzer (GeXP-PCR). Eleven sets of chimeric primers were used to initiate the PCR, and one pair of universal primers was used for the subsequent cycles of the PCR. The specificity of GeXP-PCR for each HPV type was examined with clinical samples of single type HPV infection tested previously. The sensitivity of GeXP-PCR was evaluated by performing the assay on serial 10-fold dilutions of cloned PCR products. The GeXP-PCR achieved a sensitivity of 100 copies when all of the 11 pre-mixed plasmids containing HPV targets were present. Analyses of 124 clinical specimens using the GeXP-PCR demonstrated that the GeXP-PCR assay had comparable sensitivity and specificity to those of reported multiple PCR assay and an increased detection of HPV 11 in samples with mixed infections. In conclusion, the GeXP-PCR is a fast, sensitive, and high throughput method for the detection of multiple HPV infections.
authors.affiliation: State Key Laboratory for Molecular Virology and Genetic Engineering, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
authors.forename: Meng-Jie
authors.initials: MJ
authors.lastname: Yang
authors.forename: Le
authors.initials: L
authors.lastname: Luo
authors.forename: Kai
authors.initials: K
authors.lastname: Nie
authors.forename: Miao
authors.initials: M
authors.lastname: Wang
authors.forename: Chen
authors.initials: C
authors.lastname: Zhang
authors.forename: Jin
authors.initials: J
authors.lastname: Li
authors.forename: Xue-jun
authors.initials: XJ
authors.lastname: Ma
mesh_terms: D017931:DNA Primers / Q000235:genetics; D005260:Female; D006801:Humans; D058889:Molecular Typing; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D027383:Papillomaviridae / Q000145:classification* / Q000235:genetics*; D030361:Papillomavirus Infections / Q000821:virology*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers