中国畜牧兽医2016，Vol.43Issue(7)：1688-1693,6.DOI:10.16431/j.cnki.1671-7236.2016.07.005
环介导等温扩增法检测牛奶中结核分枝杆菌条件的优化
Study on Optimization of the Conditions of Detecting Mycobacterium tuberculosis in Milk by Technique of Loop-mediated Isothermal Amplification
摘要
Abstract
To optimize the loop-mediated isothermal amplification (LAMP)method to detect My-cobacteriumtuberculosis in milk,the highly conservative 16S rRNA gene of Mycobacteriumtuber-culosis was selected as a target to design specific primers.Compared the modified method of CATB/NaCl,bacterial genome DNA extraction kit and thermal cracking method to extract the DNA of Mycobacterium tuberculosis,the best approach was chosen.Mycobacterium tuberculosis suspension liquid was diluted with sterilization milk to confirm the susceptibility of this assay. And then Brucella,Escherichia coli,Staphylococcus aureus,Listeria monocytogenes,Bacillus pasteurii and Salmonella were used for specificity detection.The results showed that the modified method of CATB/NaCl was better than the others.The sensitivity of LAMP was 3×100 CFU/mL,and the specificity was 100%.The sensitivity of detecting Mycobacterium tuberculosis in milk by LAMP was 3×101 CFU/mL.关键词
环介导等温扩增/结核分枝杆菌/牛奶Key words
LAMP/Mycobacteriumtuberculosis/milk分类
医药卫生引用本文复制引用
杨敏,王慧党,于潞,吴长新,张辉,曹旭东,陈创夫..环介导等温扩增法检测牛奶中结核分枝杆菌条件的优化[J].中国畜牧兽医,2016,43(7):1688-1693,6.基金项目
“十二五”国家科技重大专项(2013ZX10003003-002) （2013ZX10003003-002）
国家自然科学基金(31060333) （31060333）
