导流杂交基因芯片技术在人乳头状瘤病毒感染分型检测中的临床应用

Application of flow-through hybrization and gene chip on human papillomavious subtype detection in clinical

目的 探讨核酸分子快速导流杂交基因芯片技术（HybriMax）在临床人乳头状瘤病毒（human papillomavious，HPV）感染分型检测中的应用。方法采用HybriMax技术，对门诊就诊的3086例有性生活史妇女，进行下生殖道HPV感染分型检测。463例宫颈病变患者在阴道镜下定点活检病理诊断，以病理诊断为金标准评价HybriMax在发现宫颈病变中的价值，并对80例HPV16阳性的患者DNA进行E6／E7基因测序，确定其分型的准确性。结果21种HPV亚型均被检出，3086例门诊妇女中HPV阳性率63．1％（1947／3086），检出率在5．0％以上的有7种，依次为HPVl6（15．9％、490／3086）、58（11．2％、346／3086）、52（8．5％、261／3086）、33（6．3％、195／3086）、53（6．2％、192／3086）、6（5．6％、172／3086）和CP8304（5．0％、155／3086）型。HybriMax技术发现高级别鳞状上皮内瘤变（HSIL，CINII＋CINⅢ）的敏感度为95．49％（95％6＇I=95．44％～98．27％）、特异度34．85％（95％CI=32．59％～37．57％）、阳性预测值37．13％（95％CI=30．79％～40．96％），阴性预测值95．04％（95％6＇I=89．24％～98．44％）。HPV16E6／E7基因序列检测对HPV16型的准确性达100％，同时发现有型内变异存在。结论HybriMax技术检测HPV分型准确性高，对发现HSIL有较好的敏感性和特异性，是临床HPV感染分型检测的有效方法。

Objectives To evaluate the application of flow-through hybrization and gene chip （HybriMax） on human papillomavious （HPV） subtype detection in clinical. Method 3 086 sexually active females from out-patient department were selected for HPV subtype analysis using HybriMax. Cervical tissue samples were taken under the colposcope from 463 females who had cervical lesion for pathological analysis. The predictive value of HybriMax in cervical abnormality was compared with pathological results, which were used as a golden standard. DNA sequence in HPV E6/E7 region was performed among 80 females with HPV 16 positive by HybriMax to determine the accuracy of HybriMax. Results All 21 different subtypes were found and total HPV positive rate was 63.1% （ 1 947/3 086）. Among the 21 HPV subtypes, 5 of them had a infection rate over 5% and they were HPH16 （ 15.9%, 490/3 086 ）, 58 （ 11.2%, 346/3 086 ）, 52 （8.5% ,261/3 086） ,33（6. 3% ,195/3 086） ,53（6.2% ,192/3 086）,6（5.6% ,172/3 086）, and CP8304 （5.0% ,155/3 086）. The sensitivity for high degree squamous intraepithelial dysplasia（ HSIL, CIN Ⅱ ＋ CIN Ⅲ ） by HybriMax was 95.49% （95% CI = 95.44% - 98.27% ）, while specificity was 34. 85% （95% CI = 32. 59% - 37.57% ） ,Positive predictive value for HSIL was 37. 13% （95 % CI = 30. 79% - 40. 96% ）, while negative predictive vale was 95.04% （95% CI = 89. 24% -98.44% ）. Eighty sequence results in E6/ E7 region completely matched to HybriMax results. Conclusion HybriMax has a high accurate rate in HPV subtype diction with good sensitivity and specificity for HSIL and above. It is an effective method to detect HPV subtype in clinical.