多重PCR-毛细管电泳法的应用及HPV感染亚型分布特征分析

Application of multiplex PCR capillary electrophoresis and analysis of distribution characteristics of HPV infection subtypes

目的分析河北省石家庄地区人乳头状瘤病毒(human papillomavirus,HPV)感染特征以及HPV亚型分布情况,探讨多重PCR-毛细管电泳片段分析法的应用评估。方法采用横断面调查研究,2022年5至8月期间纳入河北省人民医院434例女性(年龄范围17~74岁,就诊者260例、查体者174例)进行横断面调查,应用多重PCR-毛细管电泳片段分析法进行HPV分型检测。以多重荧光定量PCR试剂盒作为参考,采用卡方检验分析多重PCR-毛细管电泳片段分析法的诊断效果,Kappa值分析其一致性。结果总HPV感染率为45.85%(199/434),其中高危型HPV(HR-HPV)阳性率为35.48%(154/434)、低危型HPV(LR-HPV)3.92%(17/434)、HR-HPV和LR-HPV混合感染6.45%(28/434)、单一型别感染率27.88%(121/434)和多型别17.97%(78/434)。HPV52(9.68%,42/434)、HPV16(6.91%,30/434)、HPV58(6.91%,30/434)为常见HPV亚型。查体者阳性率为45.40%(79/174),接近于就诊者阳性率为54.76%(120/260),差异无统计学意义(χ^(2)=0.024,P>0.05)。17~30岁年龄组感染率最高为56.32%(46/84),各年龄组之间阳性率差异无统计学意义(χ^(2)=4.123,P>0.05)。以多重荧光定量PCR为参考,多重PCR-毛细管电泳片段分析法的敏感度和特异度分别是92.96%和94.04%,Kappa值为0.870。结论HPV感染可能呈现年轻化,HR-HPV感染阳性率最高,以HPV52、16、58为主要感染亚型。多重PCR-毛细管电泳片段分析法与多重荧光定量PCR相比的检测结果具有高度一致性,适合HPV DNA分型检测。

Objective To analyze the characteristics of human papillomavirus(HPV)infection and the distribution of HPV subtypes in Shijiazhuang,Hebei Province,and to explore the application evaluation of multiple PCR capillary electrophoresis fragment analysis for HPV typing test.Methods A population-based cross-sectional study was conducted among 434 women(age range 17 to 74 years old,260 patients and 174 physical examinations)included from May to August 2022 in Hebei General Hospital.HPV typing was detected by multiple PCR-capillary electrophoresis fragment analysis.Using the multiple fluorescence quantitative PCR kit as a reference,Chi-square test was used to analyze the diagnostic effect of multiple PCR-capillary electrophoresis fragment analysis,and the consistency was analyzed by Kappa value.Results The total HPV infection rate was 45.85%(199/434),including 35.48%(154/434)of high-risk HPV(HR-HPV),3.92%(17/434)of low-risk HPV(LR-HPV),6.45%(28/434)of HR-HPV and LR-HPV mixed infection,27.88%(121/434)of single type HPV and 17.97%(78/434)of multi type HPV.HPV52(9.68%,42/434),HPV16(6.91%,30/434),and HPV58(6.91%,30/434)are common HPV subtypes.The positive rate of physical examination was 45.40%(79/174),which was slightly lower than that of patients 46.15%(120/260),there was no significant difference(χ^(2)=0.024,P>0.05).The highest infection rate in the 17-30 age group was 54.76%(46/84),and there was no statistical difference among the age groups(χ^(2)=4.123,P>0.05).The sensitivity and specificity of multiplex PCR capillary electrophoresis fragment analysis were 92.96%and 94.04%,respectively,and Kappa value was 0.870,with the multiplex fluorescent quantitative PCR as the reference.Conclusion HPV infection may appear younger,and the positive rate of HR-HPV infection is the highest,with HPV52,16,58 as the main infection subtypes.The detection results of multiplex PCR capillary electrophoresis fragment analysis method are highly consistent with those of multiplex fluorescent quantitative PCR method,which is suitable for HPV DNA typing.