Validasi Metode Real-Time Polymerase Chain Reaction (qPCR) untuk Identifikasi DNA Anjing (Canis familiaris) dalam Pangan Asal Hewan

Abstract

Konsumsi daging anjing di Indonesia banyak dikaitkan dengan etnis ataupun wilayah tertentu seperti etnis Batak Karo dan Batak Toba di Sumatera Utara serta etnis Minahasa di Sulawesi Utara. Penelitian ini bertujuan menentukan validitas metode dalam pengujian identifikasi DNA anjing pada produk pangan asal hewan. Sampel sebagai kontrol positif adalah campuran daging anjing dan daging ayam (1:9) dan daging ayam sebagai kontrol negatif. Pengujian validasi DNA anjing menggunakan real-time polymerase chain reaction (qPCR) dengan SYBYR green melting curve analysis. Parameter uji yang digunakan meliputi repeatability, reproducibility, limit of detection, dan spesificity. Hasil penelitian pada parameter repeatability menunjukkan bahwa sampel positif teramplifikasi seluruhnya, sedangkan sampel negatif tidak teramplifikasi. Pengujian nilai t terhadap nilai Ct dan Tm pada uji reproducibility tidak menunjukkan adanya perbedaan yang nyata dalam mendeteksi DNA anjing dari kedua pengulangan menggunakan real-time PCR. Batas limit deteksi DNA anjing yang didapatkan sebesar 0,03125% serta tidak terdapat cross reaction dalam pengujian terhadap spesifisitasnya. Real-Time PCR dapat dijadikan metode pengujian untuk identifikasi DNA anjing ada dalam produk pangan asal hewan.

The consumption of dog meat in Indonesia is mostly related to certain ethnicities or regions, such as the Batak Karo and Batak Toba ethnic groups in North Sumatra and the Minahasa ethnicity in North Sulawesi. This study aimed to determine the validity of the method in testing the identification of dog DNA in food of animal origin. Samples as positive control were the mixture of dog meat and chicken meat (1:9) and chicken meat as negative control. The examination on validation of DNA dog was conducted using real-time polymerase chain reaction (qPCR) with SYBYR green melting curve analysis. The test parameters used included repeatability, reproducibility, limit of detection, and specificity. The results of the research on repeatability parameters showed that the positive samples were all amplified, while the negative samples were not amplified. Testing the t value against the Ct and Tm values in the reproducibility test did not show any significant difference in detecting dog DNA from the two repetitions using real- time PCR. The limit for dog DNA detection was 0.03125% and there was no cross

reaction in testing for its specificity. Real-Time PCR can be used as a testing method for identifying dog DNA in food products of animal origin.