实时荧光PCR法在高危型HPV检测中的应用价值

Application value of real-time fluorescent PCR method in detection of high-risk HPV

目的分析实时荧光聚合酶链式反应(PCR)法在高危型人乳头瘤病毒(HPV)检测中的应用价值。方法选取2020年1月至12月在本院门诊就诊的200例疑似高危型HPV感染的患者作为研究对象,均采用PCR-反向点杂交法、杂交捕获Ⅱ代(HC-Ⅱ)法、实时荧光PCR法检测高危型HPV感染情况。以病理组织学检测结果为金标准,分析三种检测方法对高危型HPV的阳性检出率及诊断效能;比较三种检测方法对不同级别宫颈病变[慢性炎症、宫颈上皮内瘤变(CIN)、宫颈癌]高危型HPV的阳性检出率;比较不同宫颈病变组织的常见高危型HPV-DNA水平。结果病理组织学检测结果显示,慢性炎症60例;CINⅠ级61例,Ⅱ级44例,Ⅲ级30例;宫颈鳞状细胞癌5例。高危型HPV感染检出率为71.00%(142/200),按照检出例数由多到少依次为HPV 16型53例、18型33例、31型16例、35型9例、39型6例、45型5例、33型4例、52型4例、51型3例、56型3例、68型3例、58型2例、59型1例。实时荧光PCR法检测高危型HPV的阳性检出率、灵敏度、特异度、准确度均高于PCR-反向点杂交法和HC-Ⅱ法,漏诊率、误诊率均低于PCR-反向点杂交法和HC-Ⅱ法(P<0.05)。实时荧光PCR法与PCR-反向点杂交法、HC-Ⅱ法对不同级别宫颈病变高危型HPV的阳性检出率比较,差异无统计学意义(P>0.05)。宫颈癌患者的HPV 16、18型DNA平均拷贝数均高于CIN患者(P<0.05)。结论实时荧光PCR法在高危型HPV检测中的应用价值显著,具有较高的检出率、灵敏度、特异度、准确度,值得临床推广应用。

Objective To analyze the application value of real-time fluorescent polymerase chain reaction(PCR)method in detection of high-risk human papilloma virus(HPV).Methods A total of 200 patients with suspected highrisk HPV infection who visited the outpatient clinic of our hospital from January to December 2020 were selected as the research objects,and PCR-reverse dot blot method,hybrid capture Ⅱ(HC-Ⅱ)method,and real-time fluorescent PCR method were used to detect the infection of high-risk HPV.Taking the results of histopathological examination as the gold standard,the positive detection rate and diagnostic efficacy of the three detection methods for high-risk HPV were analyzed;the positive detection rate of the three detection methods for high-risk HPV in different grades of cervical lesions[chronic inflammation,cervical intraepithelial neoplasia(CIN),cervical carcinoma]were compared;the levels of common high-risk HPV-DNA in different cervical lesions were compared.Results The results of histopathological examination showed that there were 60 cases of chronic inflammation;61 cases of CIN grade Ⅰ,44 cases of grade Ⅱ and 30 cases of grade Ⅲ;5 cases of cervical squamous cell carcinoma.The detection rate of high-risk HPV infection was 71.00%(142/200),according to the number of detected cases from more to less,there were 53 cases of HPV type16,33 cases of HPV type 18,16 cases of HPV type 31,9 cases of HPV type 35,6 cases of HPV type 39,5 cases of HPV type 45,4 cases of HPV type 33,4 cases of HPV type 52,3 cases of HPV type 51,3 cases of HPV type 56,3cases of HPV type 68,2 cases of HPV type 58,and 1 case of HPV type 59.The positive detection rate,sensitivity,specificity and accuracy of real-time fluorescent PCR method for detection of high-risk HPV were higher than those of PCR-reverse dot blot method and HC-Ⅱ method,and the missed diagnosis rate and misdiagnosis rate were lower than those of PCR-reverse dot blot method and HC-Ⅱ method(P<0.05).There was no significant difference between realtime fluorescent PCR method and PCR-reverse dot blot method and HC-Ⅱ method in the positive detection rate of high-risk HPV in different grades of cervical lesions(P>0.05).The average copy number of HPV 16 and 18 DNA in cervical cancer patients were higher than those in CIN patients(P<0.05).Conclusion Real-time fluorescent PCR method in detection of high-risk HPV has a significant application value,it has high detection rate,sensitivity,specificity and accuracy,which is worthy of clinical promotion and application.