TDI-FP法检测宫颈癌组织中HPV16E7基因647位点突变

Detection of HPV16 E7 gene nt647 point mutation in cervical cancerous tissue using TDI-FP technique

目的人乳头瘤病毒(HPV)16型E6E7基因的变异可能和HPV的致癌能力直接相关,E7647位点的突变可能增强了致癌能力.本课题通过应用TDI-FP法检测宫颈癌HPV16阳性标本中E7647位点突变情况,可以对确定高危人群、恶性病变的预警筛查、辅助诊断、疗效判定及预后提供临床价值,并可以为宫颈癌及其癌前病变的基因治疗、疫苗治疗和预防提供分子水平方面的重要依据.方法TDI-FP技术是一种在聚合酶链反应的基础上具备液相探针杂交和碱基掺入三重特异反应的检测新方法,具有极高的特异性和敏感性.本课题实验在TDI-FP方法的基础上检测宫颈癌患者HPV16E7基因647位点突变情况,以进一步明确这些位点突变与宫颈癌变的相关性.结果应用TDI-FP法检测宫颈癌组织中HPV16型E7647位突变率为32.35%,HPV16型E7647位突变与对照组相比具有统计学差异(χ2=12.437,P<0.001).结论本实验初步提示宫颈癌患者中HPV16型E7647位突变可能是HPV致癌的一个高危因素.

AIM： To detect HPV16 E7 nt647 point mutation in cervical cancerous tissue using TDI-FP technique in order to provide valuable clinical data for warning, detection, diagnosis, gene therapy, prevention and prognosis of cervical cancer and precancerous lesion with HPV infections. METHODS： TDI-FP technique is a compounded reaction including PCR, probe hybridization and base ineorporation with high specificity and sensitivity. We detected the point mutation of HPV16 E7 nt647 in the detected HPV 16 positive cervical cancerous tissue by the TDI-FP method, which can explain the correlation between the point mutation and cervical cancer. RESULTS： The HPV16 E7 nt647 point mutation rate was 32.35% in the study. The point mutation rate of HPV16 E7 nt647 was significantly different from that of the control group （x^2 = 12. 437, P 〈 0. 001 ）. CONCLUSION ： The point mutation of HPV16 E7 nt647 may be a high risk factor for cervical cancer.