CAJ | 학술논문

为建立一种快速、敏感、特异、有效的检测霍乱弧菌方法，以有效预防及控制霍乱弧菌感染，针对霍乱弧菌 ctxA 基因的保守区域设计引物，应用实时浊度仪进行环介导等温扩增（LAMP），研究其特异性与灵敏性，并利用该体系检测海产品中的霍乱弧菌。结果显示，含 ctxA 基因的霍乱弧菌可得到特异性扩增，而其他与霍乱弧菌（含 ctxA 基因）共存于海产品中的7种细菌均未得到扩增，DNA 检测下限为81 fg/μL。用建立的实时浊度 LAMP 检测方法对150份海产品进行检测，检出含霍乱弧菌的海产品2份，与荧光定量 PCR 结果一致。表明建立的 LAMP 检测方法可用于海产品中霍乱弧菌的检测。
위건립일충쾌속、민감、특이、유효적검측곽란호균방법，이유효예방급공제곽란호균감염，침대곽란호균 ctxA 기인적보수구역설계인물，응용실시탁도의진행배개도등온확증（LAMP），연구기특이성여령민성，병이용해체계검측해산품중적곽란호균。결과현시，함 ctxA 기인적곽란호균가득도특이성확증，이기타여곽란호균（함 ctxA 기인）공존우해산품중적7충세균균미득도확증，DNA 검측하한위81 fg/μL。용건립적실시탁도 LAMP 검측방법대150빈해산품진행검측，검출함곽란호균적해산품2빈，여형광정량 PCR 결과일치。표명건립적 LAMP 검측방법가용우해산품중곽란호균적검측。
In order to establish a rapid,sensitive,specific,and effective method for diagnosis of vibrio cholera,to prevent and control the bacteria infection,a loop-mediated isothermal amplification(LAMP)assay was developed with a set of primers designed for V.cholera ctxA gene.The real-time monitoring of the LAMP reaction was achieved by real-time turbidimeter,and the specificity and sensitivity were tested.The results showed that V.cholera with ctxA gene exhibited typical curves,and 7 strains of other bacteria concurred withV.cholera in seafood.The detection limitation of this method was 81 fg/μL for purified genomic DNA.Furthermore,a total of 150 seafood samples were tested by the established method and the virulence ctxA gene was detected from 2 samples,which was accordant with the result by the real-time quantitative PCR. This assay is rapid and reliable for detection ofV.cholera.