肠炎沙门菌环介导等温扩增快速检测方法的建立  

Exploration of Loop-Mediated Isothermal Amplification for Detection of Salmonella Enteritidis

作　　者：张海艳[1] 周赞虎 胡元庆 刘斌[4] 

机构地区：[1]漳州卫生职业学院,福建漳州363000 [2]漳州出入境检验检疫局 [3]闽南师范大学生物科学与技术学院 [4]西北农林科技大学

出　　处：《检验检疫学刊》2017年第6期5-9,共5页Journal of Inspection and Quarantine

基　　金：福建省漳州市自然科学基金项目(ZZ2014J35)

摘　　要：利用环介导等温扩增技术建立肠炎沙门菌快速检测方法。根据肠炎沙门菌SEN1393基因特异序列设计引物,用凝胶电泳测试实验结果,再优选反应体系,最后检测牛乳样品中肠炎沙门菌。试验结果显示该方法能够在60℃条件下1 h内检出肠炎沙门菌,对5株沙门菌和27株非沙门菌检测,只有肠炎沙门菌可以检测出阳性扩增,所设计的引物有良好的特异性;DNA检测灵敏度为94 fg/μL。该方法具有高效、特异性强和敏感性高等特点,可满足肠炎沙门菌快速检测筛选的要求。A Loop-mediated isothermal amplification（LAMP） method was established for detection of Salmonella Enteritidis. The primers were designed according to the SEN1393 sequence of Salmonella Enteritidis,and the LAMP method was constructed and optimized based on these primers. The sensitivity and specificity of LAMP were tested in this study. The results showed the Salmonella Enteritidis could be rapidly and specifically identified by the LAMP method within 1 h at constant temperature of 60℃.Five Salmonella Enteritidis strains were able to be amplified specific fragments of gene SEN1393,while27 strains of non-Salmonella Enteritidis could not,and no false positive or false negative results occured.The sensitivity of the method was as low as 94 fg/μL. The Salmonella Enteritidis LAMP method is effective,sensitive and useful for the detection of Salmonella Enteritidis.

关 键 词：环介导等温扩增 肠炎沙门菌 敏感性与特异性 

分 类 号：TS207.4[轻工技术与工程—食品科学]