[The development and clinical application of papillomavirus genotyping by DNA chip].
OBJECTIVE
To develop a new platform for genotyping human papillomavirus (HPV) and to investigate its effect in clinical application.


METHODS
A pair of common primers of 18 HPV subtypes for PCR, was designed in HPV conservative L1 region. Genotyping probes for detecting 15 high-risk HPV subtypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66 and 68, together with 3 low-risk HPV 6, 11 and 42 were selected respectively from Genbank and fixed on membrane to make DNA chip. PCR amplification and DNA chip technology were optimized. 100 clinical samples were used to investigate the effect of HPV genotyping DNA chip. Veracity of the genotyping results was verified by sequencing.


RESULTS
From the 100 clinical samples, 30 were found to be HPV positive, including high-risk HPV subtypes 16, 18, 33, 45, 51, 58, and 66, and low-risk HPV 6, 11 and 42. The sensitivity tested by standard samples was up to 10 copies of HPV DNA.


CONCLUSIONS
The HPV genotyping system developed here with DNA chip showed high sensitivity and specificity, suitable to be applied in clinical practice for HPV diagnosis and investigation on the prevalence of HPV sub-types.
Yang
Guang
G
Institute for Clinical Medicine, The First People's Hospital of Foshan, Foshan 528000, China.
Liang
Cai-hong
CH
Cui
Jin-huan
JH
Chen
Shu
S
D015344:DNA Probes, HPV / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000821:virology
D015344:DNA Probes, HPV; D004279:DNA, Viral