[논문]Loop-Mediated Isothermal Amplification Assay Targeting the femA Gene for Rapid Detection of Staphylococcus aureus from Clinical and Food Samples

Zhao, Xihong; Li, Yanmei; Park, Myoungsu; Wang, Jun; Zhang, Youhong; He, Xiaowei; Forghani, Fereidoun; Wang, Li; Yu, Guangchao; Oh, Deog-Hwan

doi:10.4014/jmb.1207.07022

[국내논문] Loop-Mediated Isothermal Amplification Assay Targeting the femA Gene for Rapid Detection of Staphylococcus aureus from Clinical and Food Samples 원문보기

Journal of microbiology and biotechnology, v.23 no.2, 2013년, pp.246 - 250

Zhao, Xihong (Key Laboratory for Green Chemical Process of Ministry of Education, School of Chemical Engineering and Pharmacy, Wuhan Institute of Technology) , Li, Yanmei (Guangzhou Women and Children's Medical Center) , Park, Myoungsu (Department of Food Science and Biotechnology and Institute of Bioscience and Biotechnology, Kangwon National University) , Wang, Jun (Department of Food Science and Biotechnology and Institute of Bioscience and Biotechnology, Kangwon National University) , Zhang, Youhong (Key Laboratory for Green Chemical Process of Ministry of Education, School of Chemical Engineering and Pharmacy, Wuhan Institute of Technology) , He, Xiaowei (College of Light Industry and Food Sciences, South China University of Technology) , Forghani, Fereidoun (Department of Food Science and Biotechnology and Institute of Bioscience and Biotechnology, Kangwon National University) , Wang, Li (Food Safety Key Laboratory of Guangdong Province, College of Food Science, South China Agricultural University) , Yu, Guangchao (First Affiliated Hospital of Jinan University) , Oh, Deog-Hwan (Department of Food Science and Biotechnology and In)

Abstract ▼ AI-Helper

In this study, a loop-mediated isothermal amplification (LAMP) method to rapidly detect Staphylococcus aureus strains was developed and evaluated by extensively applying a large number of S. aureus isolates from clinical and food samples. Six primers were specially designed for recognizing eight distinct sequences on the species-specific femA gene of S. aureus. The detection limits were 100 fg DNA/tube and $10^4$ CFU/ml. The LAMP assay was applied to 432 S. aureus strains isolated from 118 clinical and 314 food samples. Total detection rates for the LAMP and polymerase chain reaction assays were 98.4% (306/311) and 89.4% (278/311), respectively.

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제안 방법

Heating and isothermal amplification were performed in a water bath with a heating block. Amplification products of the LAMP assay were dyed with SYBR Green I and then determined through both visual observations of the color change and an ultraviolet (UV) fluorescence assay. This experiment was performed twice to ensure reproducibility.

성능/효과

From the 432 strains, 306 and 278 were detected as positive by LAMP and PCR, respectively. Total detection rates for the LAMP and PCR assays were 98.4% (306/311) vs. 89.4% (278/311), with a 100% specificity and positive predictive value (PPV) obtained by both assays. In comparison with conventional PCR methods, LAMP yielded a better detection rate and negative predictive value (NPV), whereas both assays achieved a high PPV.
(B) Intensely bright ones are determined to be positive and those lacking appreciable fluorescence are deemed negative.
4%. In conclusion, the rapidity, ease of use, cost-effectiveness, broadness, and flexibility of applying the present LAMP assay will aid in the broad molecular detection of S. aureus in both clinical and food samples.

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