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Genotyping of 11 human papillomaviruses by multiplex PCR with a GeXP analyzer
Author(s) -
Yang MengJie,
Luo Le,
Nie Kai,
Wang Miao,
Zhang Chen,
Li Jin,
Ma Xuejun
Publication year - 2012
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.23275
Subject(s) - genotyping , multiplex polymerase chain reaction , biology , virology , multiplex , serial dilution , genotype , microbiology and biotechnology , polymerase chain reaction , plasmid , human papillomavirus , gene , genetics , medicine , pathology , alternative medicine
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. J. Med. Virol. 84:957–963, 2012. © 2012 Wiley Periodicals, Inc.