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Loop‐Mediated Isothermal Amplification Assays for Detecting Yersinia pseudotuberculosis in Milk Powders
Author(s) -
Zhang Hongwei,
Feng Jinsong,
Xue Rui,
Du XinJun,
Lu Xiaonan,
Wang Shuo
Publication year - 2014
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/1750-3841.12436
Subject(s) - loop mediated isothermal amplification , yersinia pseudotuberculosis , corynebacterium pseudotuberculosis , microbiology and biotechnology , yersinia enterocolitica , polymerase chain reaction , detection limit , genomic dna , biology , bacteria , chemistry , dna , gene , chromatography , genetics , virulence
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 Application Rapid and accurate detection of Yersinia pseudotuberculosis in milk products.