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KMID : 0545120190290101543
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Journal of Microbiology and Biotechnology
2019 Volume.29 No. 10 p.1543 ~ p.1552
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Establishment and Application of Polymerase Spiral Reaction Amplification for Salmonella Detection in Food
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Xu Wenli
Gao Jun
Zheng Haoyue
Yuan Chaowen
Hou Jinlong
Zhang Liguo
Wang Guoqing
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Abstract
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Salmonella is a common zoonotic and foodborne pathogen that causes high morbidity and mortality in developing countries. In this study, we established and validated a polymerase spiral reaction (PSR) assay which targeted the conserved invasion gene (invA) of Salmonella by SYBR Green I indicator methods. Subsequently, assays for determination of the optimal conditions for optimal specificity and sensitivity of PSR were performed. We performed comprehensive evaluations using loop-mediated isothermal amplification (LAMP) and realtime PCR. A total number of 532 samples of daily food were analyzed by PSR. Twenty-seven bacterial strains were tested in the specificity assay, from which positive results were obtained only for 14-Salmonella strains. However, none of the 13 non-Salmonella strains was amplified. Similarly with LAMP and real-time PCR, the detection limit of the PSR assay was 50 CFU/ml. The PSR method was also successfully applied to evaluate the contamination with Salmonella in 532 samples of daily food, corroborating traditional culture method data. The novel PSR method is simple, sensitive, and rapid and provides new insights into the prevention and detection of foodborne diseases.
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KEYWORD
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Salmonella, invasion gene A, isothermal amplification, rapid detection, food samples
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