目的 鉴定和评估pcr联合限制性片段长度多态性(rflp)及基因测序技术对发生于外生殖器或肛周部位的5种皮肤病与性病进行hpv dna检测和分型的可行性.方法 应用hpv通用引物对(my09/11)检测组织中hpv dna,并对hpv dna阳性片段进行纯化和回收.利用4种限制性内切酶对hpv dna阳性的pcr产物进行酶切,聚丙烯酰胺凝胶电泳(page)对酶切产物进行分型,然后用pcr直接测序法验证分型结果并对难以分辨或少见图形进行检测.结果 50份经病理确诊的临床样本中,共有35份hpv dna阳性,其中26份来自尖锐湿疣患者,8份来自鲍温样丘疹病患者,1份来自鳞状细胞癌患者.hpv dna阳性样本中,hpv6型19份,hpv11型3份,hpv16型8份,hpv6合并hpv 11型4份,hpv62型1份.测序结果与pcr-rflp判读的hpv型别相符.结论 pcr-rflp方法可用于hpv dna检测与分型.
abstract：
objective to assess polymerase chain reaction(pcr)combined with restriction fragment length polymorphism(rflp)and gene sequencing technologies in the detection and typing of hpv dna.methods tissue specimens were collected from skin diseases and venereal disease in perianal or genitals.pcr was performed with hpv dna general primers(my09/11)in tissue samples. positive fragments of hpv dna were purified and digested by restriction enzymes.the digested fragments were typed by po]yacrylamide gel electrophoresis(page).the results were verified by direct sequencing.results in 50 clinical samples there were 35 hpv dna positive,including 26 from patients with condyloma acuminatum,8 from patients with bowenoid papulosis,and 1 from patients with squamous cell carcinoma.in hpv dna positive samples,19 were hpv6,3 were hpv11,8 were hpv16,4 were hpv6 and hpv 11,and i was hpv62.sequencing results were in accordance with the pcr-rflp results .conclusion pcrrflp method is effective in the detection and typing of hpv dna.
