Rapid Detection of Live/Dead Salmonella via EMA-LAMP Method

doi:10.7506/spkx1002-6300-200922077

Abstract

Abstract:

A combinatorial method, designated as ethidium monoazide (EMA)-loop-mediated isothermal amplification (LAMP) method, was established to detect live/dead Salmonella due to the selective penetration of EMA into dead cells in the presence of live cells. DNA covalently bound to EMA can not be amplified by LAMP. In this detect system, a series of primers were specially designed to recognize six distinct sequences on target invA gene in Salmonella. Totally 241 bp target DNA was amplified and visualized as ladder-like bands on agarose gel within 60 min under isothermal condition of 65 ℃. This detection method was rapid, convenient and highly sensitive. The detection limit of this LAMP assay was 10-10 g DNA/tube, which exhibited 100- fold higher sensitivity than EMA-PCR method. This novel EMA-LAMP method for detecting live/dead bacteria will have great potential.

Key words: Salmonella, rapid detection, EMA-LAMP

CLC Number:

TS207.4

LU Yu-xia，GUO Si-yuan，SHI Lei*，LI Lin. Rapid Detection of Live/Dead Salmonella via EMA-LAMP Method[J]. FOOD SCIENCE, 2009, 30(22): 324-327.

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Rapid Detection of Live/Dead Salmonella via EMA-LAMP Method

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