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| To develop a loop mediated isothermal amplification assay for quick detection of Listeria monocytogenes |
| YAO Dong, ZHANG Ru-sheng, OU Xin-hua,et al |
| Department of Microbiological Test，Changsha Center for Disease Control and Prevention, Changsha Hunan 410001， China |
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Abstract Objective: To establish a loopmediated isothermal amplification(LAMP) assay for the rapid and specific detection of Listeria monocytogenes. Methods: Four primers regions on the hlyA gene of Listeria monocytogenes were designed and used for LAMP assay. Listeria monocytogenes DNA was amplified under isothermal conditions(65℃)for 60 min in water bath, then the amplified product was judged by naked eye, SYBR Green l staining, and electrophoresis analysis. To evaluate the specificity of the assay, 1 strain of Listeria monocytogenes and 10 strains of noneListeria 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 analysis were able to detect the products in the LAMP assay，and amplification were not observed when 10 strains of nonListeria 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.
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Received: 21 June 2012
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2012, Vol. 22 
