江苏大学学报（医学版）2012，Vol.22Issue(5)：394-397,4.
快速检测单核细胞增生李斯特菌LAMP方法的建立
To develop a loop-mediated isothermal amplification assay forquick detection of Listeria monocytogenes
摘要
Abstract
Objective: To establish a loop-mediated isothermal amplification( LAMP ) assay for the rapid and specific detection of Listeria monocy to genes. Methods: Four primers regions on the hlyA gene of Listeria monocytogenes were designed and used for LAMP assay. Listeria monocytogenes DNA was amplified under i-sothermal conditions! 65°C )for 60 min in water bath, then the amplified product was judged by naked eye, SYBR Green 1 staining, and electrophoresis analysis. To evaluate the specificity of the assay, 1 strain of Listeria monocytogenes and 10 strains of none -Listeria monocytogenes were tested by LAMP and conventional PCR assay. In addition, the detection limit of LAMP was compared with that of PCR by using the Listeria monocytogenes strain,that were serially diluted and were amplified by LAMP and PCR. Results: With one strain of Listeria monocytogenes, observation with naked eyes, SYBR Green I staining and electrophoretic a-nalysis were able to detect the products in the LAMP assay, and amplification were not observed when 10 strains of non-Listeria monocytogenes were tested. The sensitivity of LAMP was higher than that of PCR assay. The detection limit of LAMP assay for Listeria monocytogenes was 3 cfu/ml and that of PCR was > 300 cfu/ml. LAMP method was superior to conventional PCR for its rapid detection of Listeria monocytogenes, which can complete in 60 min. Conclusion: LAMP for rapid and specific detection of Listeria monocytogenes was established in this study.关键词
环介导等温扩增/单核细胞增生李斯特菌/溶血素基因Key words
loop-mediated isothermal amplification/ Listeria monocytogenes / hemolysin gene分类
医药卫生引用本文复制引用
姚栋,张如胜,欧新华,宋克云..快速检测单核细胞增生李斯特菌LAMP方法的建立[J].江苏大学学报（医学版）,2012,22(5):394-397,4.
