应用树状DNA杂交(DDH)对生殖道尖锐湿疣中HPV DNA的分型检测

从手术切除的50例生殖道尖锐湿疣新鲜标本中,以及15例正常人血清中,提取基因组DNA,同时用树状DNA杂交(dendrimer DNA hybridizalion,DDH)技术和PCR进行HPV DNA的分型检测.结果50例尖锐湿疣中,以DDH方法检测,感染HPV6型者20例,感染11型者24例,6/11型混合感染者3例,阴性3例,总检测率达94%;以PCR方法检测,HPV6型感染者21例,11型感染者24例,6/11型混合感染者3例,阴性2例,总检测率为96%.15例正常人血清中,以DDH方法检测,HPV感染的假阳性率为0%;以PCR检测,假阳性率为6.67%.还以HPV阳性标本对DDH方法做了敏感度的测定,结果阳性病例DNA检测最低浓度为97.28pg/ml.研究表明,DDH技术具有较高敏感性和高特异性,且成本较低,操作安全简便,可适用于基层中小医院较大样本量筛查.

Detection of Human Papillomavirus Genotypes in Genital Condyloma Acuminata Using Dendrimer DNA Hybridization

Human papillomaviruses(HPVs)are epithelium tropic viruses associated with several cutaneous,epithelial and mucosal lesions.Until now,more than 80 different genotypes of HPV have been identified.In this study,HPV 6 and 11 which are frequently associated with benign lesions have been detected in genital condyloma acuminata.To explore a new detection method with high sensitivity and high specificity,50 tissue samples of condyloma acuminata obtained from surgically excised specimens and 15 normal serum samples have been detected by DDH and PCR methods simultaneously.In these 50 cases with genital condyloma acuminata,using DDH method,HPV DNA was positive for 20(HPV 6),24(HPV 11)and 3(HPV 6/11),respectively,and the total positive percentage with DDH was 94%,while 21(HPV 6),24(HPV 11),3(HPV 6/11)and 96% was positive using PCR methods.In 15 normal serum samples,the false positive frequency of HPV DNA was 0/15(0%)and 1/15(6.67%)by using DDH and PCR methods respectively.In this study,we quantified the sensitivity of DNA detection of HPV positive cases by DDH method.The lowest DNA concentration of positive cases which could be detected was 97.28pg/ml.The results showed that DDH is sensitive,highly specific and low cost,and also is suitable for grass roots,small or middle sized hospitals.