Preliminary Study On The Clinical Significance Of Flow-through Hybridization Gene Chip And Real-time Quantitative PCR In Human Papilloma Virus Testing

Objective①To evaluate the accordance of flow-through hybridization gene chip and real-time quantitative PCR in human papilloma virus(HPV) testing compared with HCⅡ,and to evaluate the clinical significance for screening cervical lesions above CINⅡ;②To preliminarily estimate the distribution of different human papilloma virus genotypes in different cervical lesions;③To estimate the relationship between HPV viral load and the degrees of cervical lesions.Methods The cervical cell samples of 103 sexually active females who visited the Obstetrics and Gynaecology Hospital of Fudan University from May 2007 to Octorber 2007 were collected into this research,including 48 cases in cervicitis group,28 cases in cervical flat condyloma and low-grade CIN(CINⅠ) group and 27 cases in high-grade CIN(CINⅡ, CINⅢ)and invasive cervical cancer group.All smears were detected for HPV by flow-through hybridization gene chip,real-time quantitative PCR and Hybrid CaptureⅡ.Then,the cervical biopsies were taken through colposcopy for histology.We evaluate the accordance of HybriMax and RT-qPCR compared with HCⅡin human papllima virus(HPV) testing.The sensitivity and negative predictive value of HybriMax and RT-qPCR for cervical lesions above CINⅡwere calculated according to histology, which were considered as a golden standard.Meanwhile,we preliminarily estimated the distribution of different HPV genotypes in different cervical lesions according to HPV subtypes detection by HybriMax and tried to figure out the relationship between HPV viral load and the degrees of cervical lesions according to HPV-DNA quantitation by RT-qPCR. Statistical analysis was performed by SPSS 13.0.The x~2 test,Fisher's exact propability and Kruskal-Wallis H test were used to compare the related parameters,while P＜0.05 means the existence of significant difference in statistics.Results①The accordance of HybriMax and HCⅡfor HPV detecting were excellent,with total according rate of 86.41%and Kappa index of 0.72; and the results of RT-qPCR and HCⅡalso had good correlation,with an according rate of 96.12%and Kappa index of 0.92.The sensitivity of HybriMax for cervical lesions above CINⅡwas 96.30%,while the negative predictive value was 97.44%;and the sensitivity by RT-qPCR was 100%, while the negative predictive value was 100%.②According to the results of HPV subtypes detection by HybriMax,the most common genotypes among 103 cases were HPV 16(28.16%),HPV58(11.65%) and HPV52(7.77%), consecutively.The proportion of HPV16,31,33 subtypes in high-grade CIN and invasive cervical cancer group(74.07%,11.11%,11.11%) were higher than that in cervical flat condyloma and low-grade CIN group (28.57%,3.57%,0.00%) and also higher than that in cervicitis group(2.08%,6.25%,2.08%).However,the proportion of HPV6,11 subtypes in cervical flat condyloma and low-grade CIN group(10.71%,14.29%) were higher than that in cervicitis group(2.08%,0.00%) and also higher than that in high-grade CIN and invasive cervical cancer group(3.70%,3.70%).③Based on the HPV-DNA quantitation by RT-qPCR combined with histology, the higher the HPV viral load,the more serious the cervical lesions.Conclution①HybriMax and RT-qPCR have good accordance in HPV testing compared with HCⅡ,and also have high sensitivity and negative predictive value in screening cervical lesions above CINⅡ.Furthermore,HybriMax can detect HPV subtypes and RT-qPCR is able to quantitate HPV-DNA.However, the stabilities of them need to be promoted and some missed diagnosis did exist,so the further study with more samples is needed to verify their values.②Among all the samples,the three which prossess the highest proportion were HPV16,58,52.HPV16 was the main genotype in high-grade CIN and invasive cervical cancer,while HPV6,11 were the main genotypes in cervical flat condyloma and low-grade CIN.③The higher the HPV viral load,the more serious the cervical lesions.