Rapid Detection and Discrimination Viable Cell of Listeria monocytogenes by EMA-LAMP Assay

doi:10.3969/j.issn.1673-1689.2012.09.009

Home > Archive>Volume 31, Issue 9, 2012 >951-956. DOI:10.3969/j.issn.1673-1689.2012.09.009

Rapid Detection and Discrimination Viable Cell of Listeria monocytogenes by EMA-LAMP Assay
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                        10.3969/j.issn.1673-1689.2012.09.009
                    
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Abstract:

Viable cells of Listeria monocytogenes could be selectively discriminated from dead cells by applying Ethidium bromide monoazide(EMA) cooperated into loop-mediated isothermal amplification(LAMP) assay.The EMA-LAMP method was used for the rapid detection and identification of foodborne pathogene Listeria monocytogenes.The amplification can be obtained in an hour under the isothermal condition at 63°C by designing four LAMP specifically primers from the six different sequences on the hly gene.The EMA-LAMP assay discriminated the viable and dead cells of Listeria monocytogenes.The result showed that the optimized light exposure time to achieve cross-linking of DNA by the EMA in dead cells and to photolyse the free EMA in solution was at least 20 min,and the use of 10 μg/ml or less of EMA did not inhibit the LAMP amplification of DNA derived from viable cells,but the minimum concentration of EMA to completely inhibit the LAMP amplification of DNA derived from dead cells was 4.0 μg/ml;a detection limit of the assay for the viable cells was 20 CFU/ml.

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LU Shu-xia, XU Bin, YU Xiao-dan, JIN Xue-hua, LIN Ying. Rapid Detection and Discrimination Viable Cell of Listeria monocytogenes by EMA-LAMP Assay[J]. Journal of Food Science and Biotechnology,2012,31(9):951-956.

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