摘要
霍乱是由霍乱弧菌感染所引起的,属于甲类传染病.因此,建立一种快速、灵敏、特异的检测方法,对霍乱的预防和控制工作有十分重要的现实意义.通过霍乱弧菌外膜蛋白的ompW基因设计特异引物,建立霍乱弧菌的LAMP检测方法,同时对细菌进行扩增以检测该方法的灵敏度和特异性,并与普通PCR进行比较.实验的全部反应可在2 h内完成,且反应结果可通过肉眼观测直接判定;实验中霍乱弧菌的LAMP检测方法灵敏度是普通PCR的100倍,最低检出下限为10-5;检测霍乱弧菌均为阳性,其它13株非霍乱菌则全部阴性,特异度为100%.结果表明:该方法能快速、灵敏、特异性地检测霍乱弧菌,为快速检测病原微生物起到了很好的借鉴作用.
Cholera belong to a category of infectious diseases,which is caused by Vibrio cholerae infection.Therefore,it＇s significant to establish a rapid,sensitive and specific detection method for the prevention and control of cholera.Based on the membrane proteins gene-ompW nucleic sequence of vibrio cholerae,six pairs of primers were designed for LAMP,while the bacteria were amplified in order to detect the sensitivity and specificity of the method,and to be compared with the normal PCR experiment.All of the reaction can be performed within 2 hours,and the reaction results can be judged by visual observation;the sensitivity of LAMP was 100 times compared with conventional PCR for detection of Vibrio cholerae,and the lowest de-tection limit was 10-5;the results for detection of Vibrio cholerae were positive,while the other 13 non-cholera bacteria were all negative,meant that the specificity was 100%.The results showed that： LAMP can be quick,sensitive and specific lechnology for detection of Vibrio cholerae,which provided a good reference for the rapid detection of pathogenic microorganisms.
出处
《生命科学研究》
CAS
CSCD
北大核心
2012年第5期412-416,共5页
Life Science Research
基金
2011年湛江市科技攻关计划项目(2011C3101005)
关键词
核酸扩增技术
霍乱弧菌
检测体系
nucleic acid amplification techniques
Vibrio cholerae
Detection system
