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.


CONCLUSIONS
Rapid and accurate detection of Yersinia pseudotuberculosis in milk products.
Key Laboratory of Food Nutrition and Safety, Ministry of Education of China, Tianjin Univ. of Science and Technology, Tianjin, 300457, China; Food, Nutrition, and Health Program, Faculty of Land and Food Systems, Univ. of British Columbia, Vancouver, British Columbia, V6T 1Z4, Canada.
Hongwei
H
Zhang
Jinsong
J
Feng
Rui
R
Xue
Xin-Jun
XJ
Du
Xiaonan
X
Lu
Shuo
S
Wang
D000818:Animals; D017931:DNA Primers / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005516:Food Microbiology / Q000379:methods*; D006801:Humans; D008892:Milk / Q000382:microbiology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011208:Powders; D015011:Yersinia pseudotuberculosis / Q000235:genetics / Q000254:growth & development*
D017931:DNA Primers; D004275:DNA, Ribosomal; D011208:Powders
16S and 23S rDNA internal transcribed spacer; LAMP; Yersinia pseudotuberculosis; food safety