Sensitivity of polymerase chain reaction (PCR)-southern hybridization and conventional PCR analysis for Halal authentication of gelatin capsules

Abstract

Halal authentication of gelatin capsules was conducted using polymerase chain reaction (PCR)-southern hybridization on chip and conventional PCR analysis. The primers used in PCR-southern hybridization were targeted on mitochondria DNA (mtDNA) of cytochrome b (cyt b) gene and its amplicon was 276 bp. In conventional PCR, three pairs of mtDNA primers targeted cyt b, cytochrome oxidase II (COII) and ATP6 gene were tested, resulting of 398, 212 and 83 bp amplicons, respectively. Of 20 brands examined using PCR-southern hybridization, 6 capsules (C1–C6) were found to be porcine DNA positive but none were positive using conventional PCR method. The sensitivity of each primer in the detection of porcine DNA was 0.25 ng (cyt b), 0.1 ng (COII), 0.001 ng (Olipro™ Chip) and 0.0001 ng (ATP6). Results demonstrated that the PCR-southern hybridization on chip was useful and reliable for verifying porcine DNA in gelatin capsules compared to conventional PCR.