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Loop-mediated isothermal amplification assays for detecting Yersinia pseudotuberculosis in milk powders
Hongwei Zhang1, Jinsong Feng, Rui Xue
1Key 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.
Unlabelled:
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.
Insights
A new Loop-mediated isothermal amplification (LAMP) assay rapidly detects Yersinia pseudotuberculosis in milk powder. This sensitive method offers a faster alternative to traditional polymerase chain reaction for food safety testing.
Area of Science:
- Food Microbiology
- Molecular Diagnostics
- Bacterial Pathogenesis
Background:
- Yersinia pseudotuberculosis is a Gram-negative foodborne pathogen causing enteritis, septicemia, and reactive arthritis.
- Accurate and rapid detection is crucial for public health and food safety.
Purpose of the Study:
- To develop a sensitive and rapid Loop-mediated isothermal amplification (LAMP) assay for detecting Yersinia pseudotuberculosis.
- To target the 16S-23S rDNA internal transcribed spacer (ITS) region for enhanced specificity.
Main Methods:
- Development of a LAMP assay targeting the 16S-23S rDNA ITS region of Y. pseudotuberculosis.
- Detection of DNA amplification via visible white precipitate, completed within 1 hour.
- Evaluation of detection limits using genomic DNA and spiked milk powder samples, with and without enrichment.
Main Results:
- The LAMP assay demonstrated a detection limit of 10^0 fg/reaction for genomic DNA.
- A detection limit of 10^0 CFU/100 g milk powder was achieved with 12-hour enrichment.
- The assay proved 100 times more sensitive than conventional PCR and correctly identified 18 contaminations in 236 milk powder products.
Conclusions:
- The developed LAMP assay provides a rapid and highly sensitive method for Yersinia pseudotuberculosis detection.
- This assay facilitates efficient screening of agricultural and food products, enhancing food safety protocols.
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