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Rapid and sensitive detection of Pseudomonas aeruginosa in bottled water by loop-mediated isothermal amplification

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Abstract

A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Pseudomonas aeruginosa in bottled water was developed and evaluated. Four primers, including two outer primers and two inner primers, were specially designed by targeting the ecfX gene. The LAMP assay showed high specificity, with no false-positive or false-negative results among the 108 bacterial strains tested, including 81 strains of P. aeruginosa. The detection limit in pure culture was 15.7 CFU/mL, which is approximately 100-fold more sensitive than that of ecfX-PCR. In artificially contaminated water samples spiked with low level (3.1 CFU/250 mL) of P. aeruginosa, the LAMP assay could detect the target organisms accurately after 10 h of enrichment, in contrast to 12 h of enrichment for ecfX-PCR. In the case of bottled water samples, 9.17 % (10/109) of the samples were found to be positive by LAMP, which was in accordance with the results from GB 8538-2008 method. The LAMP assay established in this study is a simple, sensitive, and rapid protocol for the detection of P. aeruginosa. It provides an important diagnostic tool for the drinking water produce industry and regulatory agencies to better control potential risks associated with consumption of bottled water.

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Acknowledgments

Shuhong Zhang and Xiaoke Xu contributed equally to this work. This work was supported by research grants from the Science and Technology Planning Project of Guangdong Province (2010B031000020). We would like to express our gratitude to Dr Treena Burgess (Murdoch University, Australia) for constructive comments. We are also thankful to the reviewers for their valuable suggestions and comments.

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Authors and Affiliations

  1. State Key Laboratory of Applied Microbiology (Ministry—Guangdong Province Jointly Breeding Base), Guangzhou, China

    Shuhong Zhang, Xiaoke Xu, Qingping Wu & Jumei Zhang

  2. Guangdong Provincial Public Laboratory for Applied and New Technology of Microbiology, Guangzhou, China

    Shuhong Zhang, Xiaoke Xu, Qingping Wu & Jumei Zhang

  3. Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangzhou, China

    Shuhong Zhang, Xiaoke Xu, Qingping Wu & Jumei Zhang

  4. Guangdong Institute of Microbiology, Guangzhou, 510070, People’s Republic of China

    Shuhong Zhang, Xiaoke Xu, Qingping Wu & Jumei Zhang

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  1. Shuhong Zhang
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  2. Xiaoke Xu
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  3. Qingping Wu
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  4. Jumei Zhang
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Correspondence to Qingping Wu.

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Zhang, S., Xu, X., Wu, Q. et al. Rapid and sensitive detection of Pseudomonas aeruginosa in bottled water by loop-mediated isothermal amplification. Eur Food Res Technol 236, 209–215 (2013). https://doi.org/10.1007/s00217-012-1876-7

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  • DOI: https://doi.org/10.1007/s00217-012-1876-7

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