A novel, sensitive, accurate multiplex loop-mediated isothermal amplification method for detection of Salmonella spp., Shigella spp. and Staphylococcus aureus in food.

摘要

For fast and efficient reasons, a multiplex loop-mediated isothermal amplification (mLAMP) to detect foodborne pathogens was studied. Three sets of LAMP primers for 3 kinds of pathogens were designed from nuc gene of S. aureus, ipaH gene of Shigella spp. and invA gene of Salmonella spp., respectively. The DNA(s) were specifically amplified in the presence of the templates from three foodborne pathogens in a single tube. The sensitivities of the mLAMP method were shown 1000 times higher in average on the detection for the genes than those of the classical PCR methods. The sequences of mLAMP assay-positive pathogen products were 99% fidelity with the individual pathogen DNA sequence by sequencing. The large number of tests with field samples showed that the specificity of the LAMP assay was high, and no cross-reaction of the primers and the templates occurred in the same tube. The mLAMP method is further improved and simplified in the detection of the three foodborne pathogens with great specificity and sensitivity.