3种方法检测宫颈脱落细胞HPV感染结果的分析

Three ways in detection of HPV infection of cervical squamous epithelial cells

目的探讨3种方法检测宫颈脱落细胞人乳头瘤病毒(HPV)感染的内在联系及与薄层细胞学检测(TCT)结果的相关性。方法用基因捕获DNA检测法(hC2)捕获样品HPV DNA解链、杂交,检测与宫颈癌高度相关的13种HPV。用实时荧光定量PCR(Q-PCR)法提取样品HPV DNA,扩增核酸在Q-PCR定量仪上检测结果。MPHC提取样品HPV DNA,扩增核酸,将扩增后的产物加到微孔板中进行杂交,加入酶标液,用酶标阅读仪测定结果。后两种方法检测病毒亚型数量与hC2法相同。本实验将hC2法与Q-PCR匹配检测260例宫颈脱落细胞HPV感染;将hC2法与MPHC匹配检测200例宫颈脱落细胞HPV感染情况。结果3种方法在鳞状上皮异常改变组中,HPV检出阳性率均为100%;在鳞状上皮良性改变组中Q-PCR和MPHC检测HPV阳性结果绝大部分与hC2相符,只有较少部分样品结果在hC2阴性结果的范围内,灵敏度均>hC2,MPHC假阳性率占2%。结论HPV检测结果与TCT的结果相结合评价宫颈疾病、宫颈癌及癌前病变是最佳方案。

Objective To investigate the efficiency of three methods to detect HPV infection of desquamated cervical epithelial cells and the relationship between methods and TCT. Methods hC2 method was used in disconnecting the HPV DNA chain in the samples to hybrid captures, and examine the 13 types of HPV which were highly related to cervical carcinoma. Q-PCR was first used to extract HPV DNA of samples and expanded nucleotide to be examined on Q-PCR to obtain results directly. MPHC was used to extract HPV DNA and to amplify nucleotide. The products was added to micro-well plate to proceed hybridization, and enzyme label was added, and results were studied in the enzyme labelling apparatus. The results of the latter two methods were the same as hC2. hC2 matched QPCR was used to examine 260 cases of desquamated cells from HPV infection against 200 cases of desquamated cells from HPV infection, and the results were compared by using MPHC matching examination. Results 1. With three methods, HPV infection was found to be 100% in all the abnormal cervical squamous epithelial cells. 2. The sensitivity of the methods, howerer, was different. The Q-PCR and MPHC had higher positive rate than hC2. Positive HPV infection rate with Q-PCR and MPHC was the same as that of hC2 in the majority of cases, as in a few cases, it was positive with Q-PCR and MPHC but negative with hC2 alone. False positive rate was 2% in MPHC Conclusion HPV testing combined with the TCT routine is the best screening program for detecting cervical cancer and precursor of cervical careinoma.