93 Journal of Food Hygiene and Safety Available online at http://www.foodhygiene.or.kr pISSN 1229-1153 / eISSN 2465-9223 J. Food Hyg. Saf. Vol. 36, No. 1, pp. 93~99 (2021) https://doi.org/10.13103/JFHS.2021.36.1.93 Research note PCR 법을 이용한 농산물 중 Clostridium perfringens 검출을 위한 전처리법 확립 최송이 1 · 서민경 2 · 윤재현 1 · 나겐드란 라잘링감 1 · 황인준 1 · 김세리 1 * 1 농촌진흥청 국립농업과학원 유해생물팀, 2 식품의약품안전처 식품식품위해평가과 Establishment of Sample Preparation Method for PCR Detection of Clostridium perfringens from Agricultural Products Song-Yi Choi 1 , Min-Kyoung Seo 2 , Jae-Hyun Yoon 1 , Nagendran Rajalingam 1 , Injun Hwang 1 , Se-Ri Kim 1 * Microbial Safety Team, National Institute of Agricultural Sciences, Rural Development Administration, Wanju, Korea Food Safety Risk Assessment Division, Natioanal Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety, Cheongju, Korea (Received October 10, 2020/Revised November 10, 2020/Accepted November 26, 2020) ABSTRACT - 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 enterotoxin- producing Clostridium perfringens from produce by polymerase chain reaction (PCR). Each produce type was inocu- lated at concentrations of 10 , 10 , 10 , 10 , 10 , and 10 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 partic- ular, 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 opti- mum sample preparation method for detection of C. perfringens by PCR. Key words : Clostridium perfringens, Sample preparation method, Produce, Polymerase chain reaction Clostridium perfringens 는 그람 양성 포자 형성 혐기성 간균으로 설사를 유발하는 주요 식중독세균이다 . C. perfringens 는 생산되는 외독소에 따라 A 형-E 형으로 분류 할 수 있다. A 형은 주로 α-toxin 을, B 형은 α-toxin, β-toxin, ε-toxin 을, C 형은 α-toxin, β-toxin 을, D 형은 α-toxin, ε-toxin 을, E 형은 α-toxin, ι-toxin 을 분비한다 . 그리고 최근에는 사람과 동물에게 괴사성 장염을 일으키는 독소로 알려진 β2 toxin 을 생성하는 C. perfringens 도 보고되고 있다 . 이 중 식중독을 일으키는 주요한 독소형은 A형이며, A 형이 생산하는 장독소(enterotoxin) 에 의해 식중독이 발생한다 . C. perfringens 가 분비하는 장독소는 대장균의 장독소와 유 사하며 주 증상은 설사로 경과는 비교적 짧은 편이다 . 국내에서 C. perfringens 에 의한 식중독사고는 최근 5 년 간(2015-2019) 총 54 건이 발생하였으며 이로 인한 환자수 는 1,842 명이었다 . 아직까지 국내에 C. perfringens 에 오 염된 농산물로 인한 식중독 사고는 보고된 바는 없으나 농산물 중 C. perfringens 의 검출은 지속적으로 보고되고 있다 . 식약처에 따르면 C. perfringens 모니터링과 관련된 12 건의 논문 및 보고서에서 총 11,647점 시료에 대한 C. perfringens 오염률 자료를 확보하여 분석하였으며 그 결 과 C. perfringens 는 육류 및 가공품에서 0-16.7%, 채소 및 가공품에서 0-22.2%, 곡류 및 가공품에서 0-6.7%의 검출 률을 보였다고 보고하였다 . 또한 2006 년 인천시 보건환 *Correspondence to: Se-Ri Kim, Microbial Safety Team, Agro- Food Safety & Crop Protection Department, National Institute of Agricultural Sciences, Rural Development Administration, Wanju 55365, Korea Tel: +82-63-238-3395, Fax: +82-63-238-3840 E-mail: seri81@korea.kr Copyright © The Korean Society of Food Hygiene and Safety. All rights reserved. The Journal of Food Hygiene and Safety is an Open-Access journal distributed under the terms of the Creative Commons Attribution Non-Commercial License(http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution,and reproduction in any medium, provided the original work is properly cited.