多重实时荧光PCR技术检测高危型人乳头瘤病毒的临床应用分析  

Evaluation of multiplex real-time PCR for detection of high risk human papillomavirus in cytological cervical samples

作　　者：张海平[1] 穆会君[2] 谢平[2] 张滨[2] 

机构地区：[1]南京医科大学附属无锡市第二人民医院皮肤科,江苏无锡214001 [2]南京医科大学附属无锡市人民医院中心实验室,江苏无锡214023

出　　处：《国际检验医学杂志》2013年第17期2256-2258,共3页International Journal of Laboratory Medicine

摘　　要：目的探讨多重实时荧光PCR技术检测高危型人乳头瘤病毒(HPV)的效能。方法应用多重实时荧光PCR试剂检测200例妇女宫颈脱落细胞高危型HPV,以凯普13种高危型人乳头状瘤病毒核酸扩增荧光检测试剂盒为对照试剂,对两者检测结果不一致和试验试剂VIC荧光通道检测显示为HPV16或HPV18阳性的样本进行DNA测序分析,评价多重实时荧光PCR技术检测高危型HPV的效能。结果 200例妇女样本中,试验试剂和对照试剂HPV高危亚型检出阳性率分别为40%和33%,两种试剂对HPV高危亚型检出阳性率比较差异有统计学意义(χ2=33.03,P<0.01)。试验试剂与对照试剂检测结果一致者165例,两种试剂检测结果总符合率为82.50%,一致性指数Kappa值为0.651 2。对照试剂检测为阴性,而试验试剂检测结果为阳性的35例标本,测序结果显示均为HPV66型。在剔除HPV66型样本后,试验试剂与对照试剂结果完全一致,总符合率为100.00%。对照试剂检测为阳性,且试验试剂VIC荧光通道阳性的标本,测序结果显示32例为HPV16型,4例为HPV18型。结论多重实时荧光PCR试剂检测高危型HPV具有较好的准确性和可靠性,是检测高危型HPV感染的又一有效手段。Objective Evaluation of multiplex real-time PCR for detecting high risk human papillomavirus.Methods Cervical cytology specimens from 200women were detected for high-risk human papillomavirus DNA with multiplex real-time fluorescence PCR kits,the samples also were detected simultaneously with high-risk HPV DNA（13subtypes）real-time PCR diagnostic kits as control.The samples with HPV16or HPV18and with inconsistent result were analyzeed by sequencing.Results Positive rate of high-risk HPV were 40%and 33% by experimental and congtrol kits respectively.The difference between them was statistically significant（χ2=33.03,P0.01）.In the 200cases,165were accordant by experimental and congtrol kits,the accordance of the two kits was 82.50%,Kappa value was 0.651 2.The 35samples which was negative by control kits and positive by experimental kits were HPV66certified with sequencing.The accordance of the two kits was 100.00% when the samples with HPV66were rejected.There were 32HPV16and 4HPV18certified with sequencing in the samples which was positive in the Fam channel of control kit and in the VIC channel of experimental kit.Conclusion Multiplex real-time PCR kit has good accuracy and reliability for detecting high-risk HPV,it is an effective method for detecting high-risk HPV.

关 键 词：乳头状瘤病毒科 聚合酶链反应 细胞诊断学 

分 类 号：R737.33[医药卫生—肿瘤]