Development of real-time PCR method for rapid and accurate detection of Centipedes (Scolopendra mutilans) in food

开发用于快速准确检测食品中蜈蚣(Scolopendra mutilans)的实时PCR方法

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Abstract

Centipedes contain pharmacologically active compounds used as important medicinal material. However, the poisons produced by centipedes can cause human diseases; therefore, its use as a food ingredient is prohibited. This is the first report to develop a real-time PCR method for detection of centipedes. The primer and probe targeting the mitochondrial cytochrome c oxidase subunit 1 (COI) gene were newly designed. The specificity was verified using ten species and was confirmed to amplify only the centipede species. The real-time PCR method exhibited good linearity with a high-determination coefficient (R (2) = 0.999) and a detection limit was 0.001 ng. The performance of our method was also verified using five real-time PCR platforms under Universal and Fast PCR conditions. Finally, its applicability to processed food was evaluated using binary insect mixtures, and at least 0.1% of centipedes was detected. Therefore, our method can specifically and sensitively detect centipedes in food, contributing to food safety.

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