中华临床感染病杂志
中華臨床感染病雜誌
중화림상감염병잡지
CHINESE JOURNAL OF CLINICAL INFECTIOUS DISEASES
2011年
2期
71-74
,共4页
柯吴坚%车雅敏%刘原君%姚卫锋%胡建中%张乃勤%杨玉民%亓玉青%孙晨薇%张俊艳
柯吳堅%車雅敏%劉原君%姚衛鋒%鬍建中%張迺勤%楊玉民%亓玉青%孫晨薇%張俊豔
가오견%차아민%류원군%요위봉%호건중%장내근%양옥민%기옥청%손신미%장준염
乳头状瘤病毒感染%聚合酶链反应%多态性,限制性片段长度%基因测序%分型
乳頭狀瘤病毒感染%聚閤酶鏈反應%多態性,限製性片段長度%基因測序%分型
유두상류병독감염%취합매련반응%다태성,한제성편단장도%기인측서%분형
Papillomavirus infections%Polymerase chain reaction%Polymorphism,restriction fragment length%Gene sequencing%Typing
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.
目的 鑒定和評估PCR聯閤限製性片段長度多態性(RFLP)及基因測序技術對髮生于外生殖器或肛週部位的5種皮膚病與性病進行HPV DNA檢測和分型的可行性.方法 應用HPV通用引物對(MY09/11)檢測組織中HPV DNA,併對HPV DNA暘性片段進行純化和迴收.利用4種限製性內切酶對HPV DNA暘性的PCR產物進行酶切,聚丙烯酰胺凝膠電泳(PAGE)對酶切產物進行分型,然後用PCR直接測序法驗證分型結果併對難以分辨或少見圖形進行檢測.結果 50份經病理確診的臨床樣本中,共有35份HPV DNA暘性,其中26份來自尖銳濕疣患者,8份來自鮑溫樣丘疹病患者,1份來自鱗狀細胞癌患者.HPV DNA暘性樣本中,HPV6型19份,HPV11型3份,HPV16型8份,HPV6閤併HPV 11型4份,HPV62型1份.測序結果與PCR-RFLP判讀的HPV型彆相符.結論 pCR-RFLP方法可用于HPV DNA檢測與分型.
목적 감정화평고PCR연합한제성편단장도다태성(RFLP)급기인측서기술대발생우외생식기혹항주부위적5충피부병여성병진행HPV DNA검측화분형적가행성.방법 응용HPV통용인물대(MY09/11)검측조직중HPV DNA,병대HPV DNA양성편단진행순화화회수.이용4충한제성내절매대HPV DNA양성적PCR산물진행매절,취병희선알응효전영(PAGE)대매절산물진행분형,연후용PCR직접측서법험증분형결과병대난이분변혹소견도형진행검측.결과 50빈경병리학진적림상양본중,공유35빈HPV DNA양성,기중26빈래자첨예습우환자,8빈래자포온양구진병환자,1빈래자린상세포암환자.HPV DNA양성양본중,HPV6형19빈,HPV11형3빈,HPV16형8빈,HPV6합병HPV 11형4빈,HPV62형1빈.측서결과여PCR-RFLP판독적HPV형별상부.결론 pCR-RFLP방법가용우HPV DNA검측여분형.
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.
