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KMID : 0881720210360010093
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Journal of Food Hygiene and Safety
2021 Volume.36 No. 1 p.93 ~ p.99
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Establishment of Sample Preparation Method for PCR Detection of Clostridium perfringens from Agricultural Products
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Choi Song-Yi
Seo Min-Kyoung
Yoon Jae-Hyun
Nagendran Rajalingam
Hwang In-Jun
Kim Se-Ri
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Abstract
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This study was undertaken to compare the efficacy of different sample preparation (stomaching, pulsifying, and sonication) and DNA extraction methods (boiling and commercial kit) for detection of enterotoxinproducing Clostridium perfringens from produce by polymerase chain reaction (PCR). Each produce type was inoculated at concentrations of 102, 103, 104, 105, 106, and 107 spores/g. Produce inoculated with spores was treated with three sample preparation methods, and DNA was extracted by boiling method and a commercial kit, followed by PCR.
The detection limit of stomached samples was lower than that of pummeled and sonicated samples by 10-100 times.
Moreover, the DNA extraction efficiency of the commercial kit was found to be superior to that of boiling. In particular, the PCR efficiency of cherry tomato and perilla leaf samples was greatly affected by sample preparation and DNA extraction method. These data suggest that DNA extraction with a commercial kit after pulsification is an optimum sample preparation method for detection of C. perfringens by PCR.
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KEYWORD
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Clostridium perfringens, Sample preparation method, Produce, Polymerase chain reaction
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