A new tool for quality control to monitor contamination of six non-halal meats in food industry by multiplex high-resolution melting analysis (HRMA)

The demand for halal food is increasing with the growing Muslim population worldwide. The aim of this research was to develop a multiplex high-resolution melting analysis (HRMA) for detecting contamination of halal foods with six non-halal meats (from donkey, cat, pig, rat, dog, and monkey). The primers designed for the analysis showed high specificity for target animal DNA and 100% accuracy. The melting profiles of the targeted animal DNA were unique and could be easily observed for species discrimination. The limit of detection for the target animal DNA was 0.01 ng, except for pig (0.001 ng). The accuracy of detection was 100% in a test to check the reliability of the developed technique using beef meatballs contaminated with 10% or 1% non-halal animal meats. Moreover, the technique was further verified using 260 commercial food products in Thailand. Two samples were found to be contaminated with pig DNA. This multiplex HRMA could be used as a rapid and high throughput technique for detecting non-halal animal contaminants in halal foods and as a useful tool for monitoring and assuring food quality for Muslim and general consumers.