环介导等温扩增法检测牛奶中结核分枝杆菌条件的优化

doi:10.16431/j.cnki.1671-7236.2016.07.005

Abstract

Abstract:

To optimize the loop-mediated isothermal amplification (LAMP) method to detect Mycobacterium tuberculosis in milk,the highly conservative 16S rRNA gene of Mycobacterium tuberculosis 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.

Key words: LAMP; Mycobacterium tuberculosis; milk

CLC Number:

R378.91⁺1

YANG Min, WANG Hui-dang, YU Lu, WU Chang-xin, Zhang Hui, CAO Xu-dong, CHEN Chuang-fu. Study on Optimization of the Conditions of Detecting Mycobacterium tuberculosis in Milk by Technique of Loop-mediated Isothermal Amplification[J]. , 2016, 43(7): 1688-1693.

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Study on Optimization of the Conditions of Detecting Mycobacterium tuberculosis in Milk by Technique of Loop-mediated Isothermal Amplification

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