CAJ | 학술논문

目的：建立一种检测志贺菌属快速敏感的荧光定量PCR方法．方法：根据志贺菌属侵袭性质粒抗原H （ipaH）编码基因序列，设计1对PCR引物特异性靶基因片段，通过检测3株志贺菌属菌株和9株非志贺菌属菌株来评价该方法的特异性；对福氏志贺菌菌液做一系列10倍稀释后进行荧光定量PCR分析敏感性，PCR 扩增产物经电泳和测序鉴定．结果：3株志贺菌属菌株出现特定大小的目的条带，9株非志贺菌属菌株未出现目的条带；测序也证实了PCR产物的特异性；荧光定量PCR检测福氏志贺菌ipaH基因的下限为200拷贝/mL．结论：本研究建立的方法快速、特异、敏感．
목적：건립일충검측지하균속쾌속민감적형광정량PCR방법．방법：근거지하균속침습성질립항원H （ipaH）편마기인서렬，설계1대PCR인물특이성파기인편단，통과검측3주지하균속균주화9주비지하균속균주래평개해방법적특이성；대복씨지하균균액주일계렬10배희석후진행형광정량PCR분석민감성，PCR 확증산물경전영화측서감정．결과：3주지하균속균주출현특정대소적목적조대，9주비지하균속균주미출현목적조대；측서야증실료PCR산물적특이성；형광정량PCR검측복씨지하균ipaH기인적하한위200고패/mL．결론：본연구건립적방법쾌속、특이、민감．
AIM:To establish a fluorescence quantitative poly-merase chain reaction (PCR)method testing shigella rapidly and sensitively.METHODS:According to shigella invaded plasmid antigen H (invasive plasmid,ipaH)coding gene sequence,a pair of PCR primers specific target gene fragment was designed and 3 strains of shigella strains and 9 strains non-shigella strains were detected to evaluate the specificity of the method,shigella flexneri bacteria liquid was done a series of ten times dilution, then analysising the sensitivity by fluorescence quantitative PCR and appraisaling the PCR amplification products by electrophoresis and sequencing.RESULTS:3 strains of shigella strains ap-peared the specific size target band,and 9 strains non-shigella strains appeared no target band;sequencing also confirmed the specificity of the PCR products;lower limit was 200 copies/mL of testing shigella ipaH gene by fluorescence quantitative PCR mothed.CONCLUSION:The method established in the study is rapid,specific and sensitive.