Molecular identification of indonesian-origin Channa striata using multiplex-PCR

Abstract

Striatin is a bioactive protein fraction extracted from Channa striata fillet. The identity of raw material of C. striata is a critical issue that must be confirmed for the consistency of its quality. PCR-based identification technique is prospective to be used to get accurate identity of C. striata. Till date, there are no specific primers been developed to identify fillet of C. striata. In this study, we have developed a specific pair of primer for identification of C. striata, namely CSTR primers. The CSTR primers were designed based on cytochrome c oxidase subunit 1 (COI) gene sequence. They were combined with beta actin (BAM) primers as internal control. The PCR assay produced two amplicons i.e. CSTR product of 269 base pairs (bp) and BAM product (138 bp) at the optimum annealing temperature (Ta) of 50°C. Seven samples of C. striata from several areas in Indonesia were positively amplified by those primers, while others popular freshwater fishes in Indonesia (nile tilapia, common carp, catfish, mozambique tilapia, pangasius and giant gouramy) and a brackish water fish (milkfish) as negative controls only produced 138 bp DNA fragment. It indicates that the CSTR primer pair detects specifically C. striata. Therefore, PCR technique using these specific primers is potentially applied for a routine analysis of C. striata for Striatin in the production process.