Development and validation of TaqMan qPCR assay for porcine DNA detection in gelatine-based foods for halal authentication

Abstract

Porcine-derived gelatine in food products is a critical concern for halal and kosher consumers, necessitating accurate and reliable detection methods. The present work aimed to develop and validate a TaqMan probe-based real-time PCR (qPCR) assay for detecting porcine DNA in gelatine-based ice creams, cakes, and pastries. After careful optimisation of the qPCR system, DNA was extracted using commercial kits, and analysed for specificity, sensitivity, and stability under various processing conditions. The assay demonstrated high sensitivity with a detection limit of 0.05 ng/µL, and confirmed the presence of porcine DNA with 99 - 100% sequence similarity. A pilot survey of 25 halal-branded commercial samples revealed no detectable porcine DNA. The present work demonstrated qPCR assay as a reliable method for routine halal authentication of gelatine-based food products, ensuring compliance with dietary regulations.