Detection of pork meat adulteration in processed meat products is a great concern in related modern food inspection techniques to Halal meat concept according to the implementation of very strict procedures for Halal food labeling. Therefore, highly specific, sensitive and easy analytical species-specific PCR procedure for processed meat adulteration detection by pork meat in both raw and cooked manufactured beef burger in laboratory followed by QIAxcel system or/and conventional agarose analysis. To achieve this PCR procedure, a laboratory experiment was designed by incorporating the pork meat in beef burger at different level of substitution ranged from 0.01 to 25% pork with manufacturing of either 100% beef or pork burger as negative and positive control, respectively. Subsequently, manufactured beef burger samples were divided into two groups included raw beef burger and fried beef burger for 5 min. then both subjected to DNA analysis. Result manifested that PCR amplification of D-Loop gene primer (186 bp) was successfully detected pork meat in both raw and cooked beef burger. While no DNA fragments were detected when the PCR procedure was applied to the 100% beef burger confirmed the primers specificity. Otherwise, for internal control, a 141bp DNA fragment of Eukaryotic 18SrRNA gene was amplified for raw and cooked beef burger DNA template. The obtained results recorded that the procedure was able to detect as low as 0.01% pork meat in beef burger. Although PCR followed by either QIAxcel or agarose techniques were suitable for target DNA fragment differentiation. However for proficiency, adequacy and performance, applying of species-specific PCR followed by the QIAxcel analysis was sensitive enough to detect 0.01% (pork/meat: w/w). In conclusion, the optimized PCR protocol followed by QIAxcel is less time consuming, provided the advantage of electronic documentation, eliminates exposure to mutagenic reagents and reducing manual errors. Interestingly, it can be suggested that optimized PCR in this research might be a rapid and sensitive method for the routine pork meat detection and quantification in raw or processed meat products e.g. beef burger. |
