摘要
利用环介导等温扩增技术建立一种快速检测水产品中霍乱弧菌的方法。针对霍乱弧菌ctxA基因的保守区域设计特异性引物,利用钙黄绿素建立可视化环介导等温扩增检测体系,研究其特异性与灵敏性。结果表明,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81fg/μL。用建立的环介导等温扩增方法对150份水产品进行检测,检出2份阳性样本,与PCR结果一致。该方法对实验仪器和操作人员的要求低,具有良好的实用性,可用于水产品中霍乱弧菌的检测。
In order to rapidly detect vibrio cholera in aquatic product,a visual loop-mediated isothermal amplification(LAMP) assay was developed with calcein/Mn^(2+).A set of primers was selected and the specificity and sensitivity was tested.The results showed that V.cholera with ctxA gene was amplified with typical color,and 7 strains of other bacteria concurred with V.cholera was not amplified.The detection limitation of this method was 81 fg/μL for purified genomic DNA.Furthermore,a total of 150 samples were tested by the established method and the virulence ctxA gene was detected from 2 samples,which was accordant with the PCR.This assay was lowered requirement for instrument and operator,suggesting good practical applicability,and suitable for detecting vibrio cholera in aquatic product.
出处
《检验检疫学刊》
2015年第3期44-47,共4页
Journal of Inspection and Quarantine
基金
浙江出入境检验检疫局科技计划项目(ZK201337)
