Loop-Mediated Isothermal Amplification Assays for Detecting Yersinia pseudotuberculosis in Milk Powders

Yersinia pseudotuberculosis is a Gram-negative foodborne pathogen that causes several diseases, such as enteritis, septicemia, and reactive arthritis. Loop-mediated isothermal amplification (LAMP) assay targeting the 16S-23S rDNA internal transcribed spacer (ITS) region was developed to detect Y. pseudotuberculosis in milk powder. The DNA amplification could be completed in 1 h, and detected by produced white precipitate visible to naked eyes. The detection limit of LAMP assay was 10(0) fg/reaction for genomic DNA, and 10(0) CFU/100 g milk powder coupled with 12 h enrichment. LAMP assay is 100 times more sensitive than conventional polymerase chain reaction method for detecting Y. pseudotuberculosis, and correctly identified 18 cases of Y. pseudotuberculosis contaminations from 236 commercial milk powder products. In conclusion, the developed LAMP assay may facilitate rapid detection of Y. pseudotuberculosis contaminations in agricultural and food products. Practical ApplicationRapid and accurate detection of Yersinia pseudotuberculosis in milk products.