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KMID : 0545120100200030569
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Journal of Microbiology and Biotechnology
2010 Volume.20 No. 3 p.569 ~ p.573
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Rapid quantification of Salmonella in seafood by real-time PCR assay
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Kumar Rakesh
Surendran P.K.
Thampuran Nirmala
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Abstract
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A quantitative detection method for Salmonella in seafood was developed using SYBR Green based real-time PCR assay. The assay was developed with pure Salmonella DNA at different dilution levels i.e 1000 to 2 gemone equivalents (GE). The sensitivity of the real-time assay for Salmonella in seeded seafood samples was determined. Minimum detection level of the real-time assay was 20 cfu/g, whereas, 2 cfu/ml was obtained for pure culture in water with the efficiency of ≥85%. The real-time assay was evaluated in repeated experiments in seeded seafood samples and regression coefficient (R2) values were calculated. The performance of the real-time assay was further assessed with the naturally contaminated seafood samples. Results highlighted that 4 out of 9 seafood sample found to be positive for Salmonella and harboured cells <100 GE/g, which were not detected by direct plating on Salmonella Chromagar media. The method developed here would be useful for rapid quantification of Salmonella in seafood and assay would be completed within 2-3 h. The ability to detect low number of Salmonella cells in seafood, present method can be used for the generation of quantitative data on Salmonella in seafood so as to implement the control measures for Salmonella contamination in seafood at harvest and post-harvest levels.
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KEYWORD
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Salmonella, Seafood, Quantitative detection, real-time PCR
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