荧光偏振方法快速检测HPV16 E7第29位密码子点突变  

Rapid detection of HPV16 E7 point mutation at codon 29 with fluorescence polarization

作　　者：陈中灿[1] 张菊[1] 高艳娥[2] 夏琳[1] 白玉杰[1] 阎小君[1] 

机构地区：[1]第四军医大学基因诊断技术研究所,陕西西安710033 [2]西安交通大学第二医院妇产科,陕西西安710004

出　　处：《第四军医大学学报》2004年第15期1403-1406,共4页Journal of the Fourth Military Medical University

摘　　要：目的 :通过TDI FP方法检测宫颈病变组织中HPV1 6E7基因第 2 9位密码子突变 (A→G)并探讨其与宫颈癌的相关性 ,同时建立一种HPV点突变检测的新方法 .方法 :以 5 3例HPV1 6阳性宫颈组织为研究对象 .首先PCR扩增含待检测位点的靶基因片段 ,然后用TDI FP方法检测荧光素碱基的掺入情况 ,判断所研究位点的基因型 .结果 :宫颈浸润癌组织中HPV1 6E7第 2 9位密码子的突变率为 4 3.4 8%(1 0 / 2 3) ,对照组为 1 0 .0 0 % (2 / 2 0 ) ,病例组为 39.39% (1 3/33) ,组间突变率差异有显著性 (P <0 .0 5 ) .该法与随机抽查标本测序结果一致率为 1 0 0 % .结论 :本研究中HPV1 6病毒发生E7第 2 9位密码子突变的情况与其他地区相比存在差异 ;HPV点突变TDI FP检测方法是一种快速简便。AIM: To investigate the relationship between the mutation and the cervical carcinoma by detecting the nucleotide missense mutation A→G at codon 29 in E7 gene of HPV16 in the cervical carcinoma tissues and to establish a new method for detecting HPV point mutation. METHODS: Totally 53 cases were examined. HPV16 E7 was first amplified by Polymerase Chain Reaction and then TDI FP was adopted to detect the types of the labeled ddNTP, by which the genotype was determined. The results were analyzed by statistics. RESULTS: The mutation rate was 43.48% (10 of 23) in invasive cervical cancer and point mutations were found in cervical tissues from 13 of the 33 case patients (39.39%), and from 2 of the 20 control subjects (10.00%). The point mutation was found statistically significantly more often in case patients than in control subjects ( P <0.05). The coherence with sequencing result was 100%. CONCLUSION: The rate of point mutation of HPV16 E7 gene in this study displays a significant geographic difference from other areas. HPV16 with the point mutation appears to be more oncogenic and may contribute to the high incidence of cervical cancer. These observations suggest that TDI FP assay of HPV point mutation is an easy, fast and high throughput method for gene mutation detection.

关 键 词：人乳头瘤病毒 突变 宫颈癌 荧光偏振 

分 类 号：R752[医药卫生—皮肤病学与性病学]