Study On HPV Detection And Cervical Lesions Diagnosis With Different HPV DNA Detecting Methods

Objectives:To evaluate the applicable value of in situ hybridization(ISH), Flow-through hybridization and gene chip(HybriMax),Real-time fluorescent quantitative PCR(FQ-PCR),and Flow fluorescent hybridization assay in cervical human papillomavirus(HPV) detection and cervical lesions diagnosis.Methods:Four different population samples who accepted cervical cancer screening in China-Japan Friendship Hospital from January 2004 to April 2007 were selected for HPV DNA detection by ISH,HybriMax,FQ-PCR,and Flow fluorescent hybridization assay,respectively.Among them,189 formalin-fixed,paraffin-embedded specimens with abnormal cervical histological diagnosis were detected for HPV16/18 genotypes by ISH;310 cytological specimens were selected for genotyping by HybriMax for 21 types of HPV;600 cytological specimens were selected for detecting by FQ-PCR for 8 types of HPV;1012 cases were selected for genotyping by Flow fluorescent hybridization assay for 15 types of HPV.All cases were tested by Hybrid CaptureⅡ(HC-Ⅱ) for 13 types of high risk HPV simultaneously.HPV DNA positive rate,test agreement and Kappa index(KI) between HC-Ⅱand the other methods were examined.Sensitivity,specifility,positive predictive value(PPV) and negative predictive value(NPV) of the different tests for detecting CINⅡor worse were compared.Results:1.In comparison with the overall positive rate of HPV DNA,HC-Ⅱwas higher than ISH(85.71%vs 55.08%),P＜0.01;HC-Ⅱwas more sensitive(90.1%vs 56.7%,P＜0.01)but less specific(27.1%vs 45.8%,P＜0.05)than ISH for detecting CINⅡor worse.2.There was no significant difference on positive rate of the same 13 high-risk HPV types between HybriMax and HC-Ⅱ(75.5%vs 73.5%,P＞0.05),The KI for the 2 assays was 0.796,with 92.3%crude agreement.The sensitivity, specifility,PPV,NPV of the 2 tests for detecting CINⅡor worse was similar.3. Comparing the overall positive rate of HPV DNA,there was no significant difference between FQ-PCR and HC-Ⅱ(56.8%vs 64.5%,P＞0.05 ).The KI for the 2 assays was 0.722,with 86.7%crude agreement.Comparing the sensitivity,specifility,PPV,NPV of the 2 tests for detecting CINⅡor worse,there was no significant difference(P＞0.05).4.There was no significant difference in positive rate of the 13 high-risk HPV types between Flow fluorescent hybridization assay and HC-Ⅱ(49.0%vs 48.5%,P＞0.05),the KI for the 2 assays was 0.721,with 86.1%crude agreement.The sensitivity of HC-Ⅱfor detecting CINⅡor worse was higher than that of Flow fluorescent hybridization assay(88.8%vs 93.1%),but there was no significant difference(P＞0.05).The specifility,PPV and NPV of HC-Ⅱfor detecting CINⅡor worse were all higher than that of Flow fluorescent hybridization assay(P value was＜0.01,＜0.05,＜0.01,respectively).Conclusions:1.HC-Ⅱhas high clinical sensitivity and excellent detection performance for HPV detection and cervical lesions diagnosis.2.ISH is not suitable for widespread clinical application for detecting HPV because of its low sensitivity and time-consuming procedure.3.HyhriMax,FQ-PCR,and Flow fluorescent hybridization assay are highly comparable to HC-Ⅱfor detecting HPV in cervical specimens.4.PCR methods have higher analytical sensitivity,but their clinical sensitivity is slight lower or sirnilar to HC-Ⅱ;The clinical performance of either HybriMax or FQ-PCR for detecting CINⅡor worse are comparable to HC-Ⅱ,but Flow fluorescent hybridization assay is lower.5.PCR methods have advantage in HPV genotyping.FQ-PCR detects less HPV types,but it can analyse quantitatively.