【摘要】 目的:建立一种适合临床男性人乳头瘤病毒(HPV)基因亚型检测的新方法,并通过检测棉拭子采集男性尿道口分泌物来验证该方法的临床检测效果。方法:通过计算机辅助,参考经典通用引物(MY09/11)和加以改进的PGMY09/11设计23种HPV基因亚型的PCR扩增引物,根据GenBank中的HPV的型特异性序列设计及合成探针,制备可同时对18种高危型:HPV-16,18,31,33,35,39,45,51,52,53,56,58,59,66,68,73,83,MM4和5种低危型:6,11,42,43,44进行分型检测的特定纸质膜芯片,并且对112例采用棉拭子采集的男性尿道口分泌物进行检测加以验证,同时,对单一阳性标本进行测序验证,对标准品进行灵敏度检测。结果:112例男性尿道口分泌物中检测出25例HPV阳性标本,其中单一HPV亚型感染13例,多重感染12例,检测出的男性HPV基因亚型有:HPV-6,11,16,18,33,35,43,56和73型九种。灵敏度可达10个拷贝HPVDNA分子。结论:该方法适用于临床进行男性HPV感染的诊断以及为开展相应的流行病、病因学调查提供切实可行的检测手段。更多还原

【Abstract】 Objective:To develop a new method for the detection of male human papillomavirus(HPV) genotypes and to investigate its clinical application value.Methods:With computer assistance and based on the classical common primers MY09/11,modified PGMY09/11 with 23 HPV subtypes for PCR and Genbank data on HPV,we designed probes for the simultaneous detection of 18 high-risk subtypes(HPV-16,18,31,33,35,39,45,51,52,53,56,58,59,66,68,73,83 and MM4) and 5 low-risk subtypes(HPV-6,11,42,43 and 44) and fixed them to the special membrane to make a DNA chip.A total of 112 male urethral samples were collected with swabs and studied for the clinical value.Meanwhile the single subtypes of HPV positive were sequenced and the standard samples detected for their sensitivity.Results:Of the total number,25 samples were found to be HPV positive,13 single HPV infection and 12 multiple infection.Nine HPV gene subtypes were noted in the samples:6,11,16,18,33,35,43,56 and 73,with sensitivity up to 10 copies of HPV DNA.Conclusion:Human papillomavirus genotyping by the membrane DNA chip is applicable to the diagnosis of male HPV infection as well as to the related epidemic and etiological investigation.更多还原