Asymmetric GP5+/6+ PCR and hybridization with fluorescence polarization assay of 15 human papillomavirus genotypes in clinical samples.
BACKGROUND
The detection of a broad spectrum of HPV genotypes has not been used widely in cervical cancer screening due to technical difficulties and high costs.


OBJECTIVE
To develop an asymmetric GP5+/6+ PCR assay and hybridization with a fluorescence polarization (FP) assay of 15 HPV genotypes.


METHODS
HPV genes in controls and samples were amplified by an asymmetric GP5+/6+ PCR. Fifteen HPV genotypic probes labeled with fluorophores hybridized with target PCR product to identify the presence of specific HPV genotypes. The HPV genotypes in samples were verified by sequence assay.


RESULTS
The genotypes determined with the hybridization and FP assay were confirmed by sequence analysis when a monotypic infection was evaluated.


CONCLUSIONS
A simple, economical and specific HPV genotyping assay has been developed that will be adequate for cervical cancer screening programs.
Ju
Zhang
Z
Institute of Gene Diagnosis, State Key Laboratory of Cancer Biology, Fourth Military Medical University, 17 Changle West Road, Xian, Shaanxi 710032, China. jianzhong@fmmu.edu.cn
Yane
Gao
G
Ding
Li
L
Guowu
Yin
Y
Yanhai
Guo
G
Yonglan
Liu
L
D002584:Cervix Uteri / Q000821:virology; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D005260:Female; D005454:Fluorescence Polarization / Q000379:methods*; D005838:Genotype; D006801:Humans; D008403:Mass Screening / Q000379:methods; D009693:Nucleic Acid Hybridization / Q000379:methods*; D027383:Papillomaviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D004279:DNA, Viral