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

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.