Minimum Limit of Detection of Pork Fat DNA in Halal Food Mixtures Using Real-Time Polymerase chain reaction (PCR) technique

Abstract

A practical scientific approach was developed and implemented to detect raw pork meat and fat. This study aimed to determine the minimal limit of detection (LOD) of pork fat DNA in a halal food mixture using Real-time PCR. The minimum LOD was determined for 0.01% and 0.001% pork fat DNA in a mixture of halal meat. A comparative analysis was conducted on DNA isolated from raw pork meat and pork fat. DNA extraction was performed using the Qiagen DNeasy Mericon Food Kit. Meat samples, including chicken, camel, and bovine, were collected from the local market, whereas pork fat samples were obtained from customs. Pork fat was mixed at 0.1%, 0.01%, and 0.001% (w/w) concentrations in the halal meat mixture. DNA band size identification of pork meat and pork fat was conducted using gel electrophoresis, and results were validated through test repeatability. The findings confirmed that Real-time PCR could detect pork fat at a minimum concentration of 0.001%, establishing it as the LOD. This study first identifies the minimum LOD of pork fat (0.001%) in halal food mixtures. Time constraints limited the research and manual DNA extraction yielded higher DNA quantities but was more time consuming than kit-based methods.