CAJ | 학술논문

目的：采用二聚体蝎型探针技术建立一种高敏感性和特异性的检测梅毒螺旋体（T P ）的荧光定量聚合酶链反应（PCR）方法。方法根据T P特异性外膜蛋白Gpd的基因序列设计引物和荧光探针，采用基因工程技术构建可用于T P定量的标准品，优化荧光定量PCR的反应体系和反应条件，建立检测T P的二聚体蝎型探针定量PCR。采用本研究建立方法和商品化荧光定量PCR试剂盒检测40例临床标本，对比分析检测结果的统计学差异。结果成功构建了T P重组质粒标准品和T P的二聚体蝎型探针定量PCR ，该方法线性范围为101～108 copy／mL ，灵敏度为10 co py／m L ，特异性和敏感性均为100．0％；二聚体蝎型探针定量PC R对疑似梅毒病例阳性检出率显著高于目前商品化Taqman荧光定量PCR试剂盒（82．5％ vs 62．5％，P＜0．05）。结论成功建立了二聚体蝎型探针荧光定量PCR快速检测T P的方法，为临床上T P的早期诊断和防控奠定了基础。
목적：채용이취체갈형탐침기술건립일충고민감성화특이성적검측매독라선체（T P ）적형광정량취합매련반응（PCR）방법。방법근거T P특이성외막단백Gpd적기인서렬설계인물화형광탐침，채용기인공정기술구건가용우T P정량적표준품，우화형광정량PCR적반응체계화반응조건，건립검측T P적이취체갈형탐침정량PCR。채용본연구건립방법화상품화형광정량PCR시제합검측40례림상표본，대비분석검측결과적통계학차이。결과성공구건료T P중조질립표준품화T P적이취체갈형탐침정량PCR ，해방법선성범위위101～108 copy／mL ，령민도위10 co py／m L ，특이성화민감성균위100．0％；이취체갈형탐침정량PC R대의사매독병례양성검출솔현저고우목전상품화Taqman형광정량PCR시제합（82．5％ vs 62．5％，P＜0．05）。결론성공건립료이취체갈형탐침형광정량PCR쾌속검측T P적방법，위림상상T P적조기진단화방공전정료기출。
Objective To establish a sensitive and specific fluorescent quantitation PCR (FQ‐PCR) by using duplex scorpion primer for rapid detection of treponema pallidum (TP) .Methods Primer and fluorescent probe were designed based on the gene order of TP outer membrane protein Gpd .Quantitative standard preparation of TP was constructed by using gene recombination .Duplex scorpion primer FQ‐PCR for rapid detection of TP was established by optimizing the reaction system and reaction condition .Totally 40 suspected cases were detected by our FQ‐PCR and commercial FQ‐PCR respectively and the difference was analyzed .Results Duplex scorpion primer FQ‐PCR for rapid detection of TP was established successfully .The linear range of the method was 101 -108 copy/mL and the sensitivity was 10 copy/mL ,susceptibility and specificity were 100 .0% .The positive detection rate of the suspected cases by our method was sharply higher than that by commercial FQ‐PCR kit (82 .5% vs 62 .5% ,P<0 .05) .Conclu‐sion Duplex scorpion primer FQ‐PCR for rapid detection of TP was established successfully ,which could be used for early diagnosis and preventive control of TP .