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KMID : 0881720180330010050
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Journal of Food Hygiene and Safety
2018 Volume.33 No. 1 p.50 ~ p.57
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Rapid Detection for Salmonella spp. by Ultrafast Real-time PCR Assay
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Kim Seok-Hwan
Lee Yu-Si
Joo In-Sun
Kwak Hyo-Sun
Chung Gyung-Tae
Kim Soon-Han
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Abstract
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Salmonella continue to be a major cause of food poisoning worldwide. The rapid detection method of food-borne Salmonella is an important food safety tool. A real-time polymerase chain reaction (PCR) has been used as a rapid method for the detection of pathogens. It has been recently reported that NBS LabChip real-time PCR is a novel, ultrafast, and chip-type-convenient real-time PCR system. We developed the assay method based on NBS Lab-Chip real-time PCR for the rapid detection of Salmonella, which its reaction time was within 20 minutes. Two target genes (invA and stn) were selected to design target specific primers and probes. The new method was validated by checking specificity and sensitivity (limit of detection). This study included forty-two target and twenty-one non-target strains to assess the specificity. This assay was able to identify the 42 Salmonella strains correctly. The limit of detection (LOD) was 101 copies/μL in Salmonella genomes DNA, while LOD incubated for 4 hr in the inoculated sausage sample ranged from 101 CFU/g to 102 CFU/g as an inoculated cell count. The assay developed in this study could be applied for the investigation of food poisoning pathogens.
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KEYWORD
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Pathogenic Salmonella spp. , Rapid detection , Real-time PCR
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