Yang Guang, Liang Cai-hong, Cui Jin-huan, Chen Shu
Institute for Clinical Medicine, The First People's Hospital of Foshan, Foshan 528000, China.
Zhonghua Liu Xing Bing Xue Za Zhi. 2006 Jan;27(1):47-9.
To develop a new platform for genotyping human papillomavirus (HPV) and to investigate its effect in clinical application.
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.
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.
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.
开发一种新的人乳头瘤病毒（HPV）基因分型平台，并研究其在临床应用中的效果。
在HPV保守的L1区域设计一对用于PCR的18种HPV亚型通用引物。分别从Genbank中选取用于检测15种高危HPV亚型16、18、31、33、35、39、45、51、52、53、56、58、59、66和68以及3种低危HPV 6、11和42的基因分型探针，并固定在膜上制成DNA芯片。对PCR扩增和DNA芯片技术进行优化。使用100份临床样本研究HPV基因分型DNA芯片的效果。通过测序验证基因分型结果的准确性。
在100份临床样本中，发现30份HPV阳性，包括高危HPV亚型16、18、33、45、51、58和66，以及低危HPV 6、11和42。标准样本检测的灵敏度高达10拷贝的HPV DNA。
这里开发的基于DNA芯片的HPV基因分型系统具有高灵敏度和特异性，适用于临床实践中HPV诊断及HPV亚型流行情况调查。
