Detection and genotyping of human papillomavirus DNA in cervical cancer tissues with fluorescence polarization.

Yan-E Gao, Ju Zhang, Jing Wu, Zhong-Can Chen, Xiao-Jun Yan
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Abstract

To evaluate the type-specific prevalence of eight common types of human papillomavirus (HPV) in patients with cervical cancer living in Shanxi, China, with fluorescence polarization detection, crude DNA extracted from 137 samples of early-stage cervical cancer (within stage IIa) and chronic cervicitis was subjected to HPV L1 consensus GP5+/GP6+ system. Then, the HPV-positive products identified by GP5 + /GP6+ PCR were genotyped based on template-directed dye-terminator incorporation assay with fluorescence polarization detection (TDI-FP): the PCR products were respectively hybridized with designed type-specific probes within the GP5+/GP6+ amplicons for eight common HPV types (HPV 6, 11, 18, 16, 31, 33, 35, and 58), and specific fluorescence-labeled ddNTPs (TAMRA-ddTTP or R110-ddGTP) were directly incorporated to the ends of the corresponding probes under directing of the corresponding template in PCR products, which was reflected and read by high FP values for TAMRA or R110. HPV DNA was detected in 38.89% (28/72) cases of chronic cervicitis, and 87.69% (57/65) cases of cervical cancer. There was a significant difference in HPV prevalence between these two groups. The four commonly identified types in patients with cervical cancer were HPV 16 (45.6%), HPV 18 (22.8%), HPV 58 (17.5%), and HPV 31 (7.02%), and in those with chronic cervicitis were HPV 16 (35.7%), HPV 11 (32.1%), HPV 6 (21.4%), and HPV 18 (10.7%). 57.14% of HPV types detected in patients with chronic cervicitis were high-risk types. HPV 16 was the most common viral type identified in both groups. Type-specific prevalence of HPV DNA has some characteristics in patients with chronic cervicitis and cervical cancer living in Shanxi, China and the worldwide uncommon type HPV 58 is relatively common in both kinds of cases. The high prevalence of HPV 58 in Chinese women should been considered in diagnosis and vaccine designs of HPV.

荧光偏振法检测宫颈癌组织中人乳头瘤病毒DNA并分型。
为评价中国山西省宫颈癌患者中8种常见型人乳头瘤病毒(HPV)的型别特异性患病率,采用荧光偏振检测方法,对137例早期宫颈癌(IIa期内)和慢性宫颈炎患者进行HPV L1共识GP5+/GP6+系统检测。然后,采用模板定向染料终止物结合荧光偏振检测(TDI-FP)技术对GP5 + /GP6+ PCR鉴定的hpv阳性产物进行基因分型;PCR产物分别与设计的八种常见HPV型(HPV 6、11、18、16、31、33、35和58)的GP5+/GP6+扩增子内的类型特异性探针杂交,在PCR产物中相应模板的指导下,将荧光标记的特异性ddNTPs (TAMRA- ddttp或R110- ddgtp)直接结合到相应探针的末端,TAMRA或R110的高FP值反映和读取。38.89%(28/72)的慢性宫颈炎患者和87.69%(57/65)的宫颈癌患者检出HPV DNA。两组间HPV患病率有显著差异。宫颈癌患者中常见的4种类型为HPV 16(45.6%)、HPV 18(22.8%)、HPV 58(17.5%)和HPV 31(7.02%),慢性宫颈炎患者中常见的4种类型为HPV 16(35.7%)、HPV 11(32.1%)、HPV 6(21.4%)和HPV 18(10.7%)。慢性宫颈炎患者检出的HPV类型中高危型占57.14%。HPV 16是两组中最常见的病毒类型。中国山西慢性宫颈炎和宫颈癌患者的HPV DNA型特异性患病率具有一定的特点,世界范围内不常见的HPV 58型在这两种病例中都相对常见。在HPV的诊断和疫苗设计中应考虑到中国女性HPV 58的高患病率。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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