Food Science Department, Faculty of Agriculture, Benha University, Moshtohor, 13736, Qaliuobia, Egypt,
Appl Microbiol Biotechnol. 2014 Dec;98(23):9805-16. doi: 10.1007/s00253-014-6084-x. Epub 2014 Oct 17.
Detection of pork meat adulteration in "halal" meat products is a crucial issue in the fields of modern food inspection according to implementation of very strict procedures for halal food labelling. Present study aims at detecting and quantifying pork adulteration in both raw and cooked manufactured sausages. This is by applying an optimized species-specific PCR procedure followed by QIAxcel capillary electrophoresis system. Manufacturing experiment was designed by incorporating pork with beef meat at 0.01 to 10 % substitution levels beside beef and pork sausages as negative and positive controls, respectively. Subsequently, sausages were divided into raw and cooked sausages then subjected to DNA extraction. Results indicated that PCR amplifications of mitochondrial D-loop and cytochrome b (cytb) genes by porcine-specific primers produced 185 and 117 bp pork-specific DNA fragments in sausages, respectively. No DNA fragments were detected when PCR was applied on beef sausage DNA confirming primers specificity. For internal control, a 141-bp DNA fragment of eukaryotic 18S ribosomal RNA (rRNA) gene was amplified from pork and beef DNA templates. Although PCR followed by either QIAxcel or agarose techniques were efficient for targeted DNA fragments differentiation even as low as 0.01 % (pork/meat: w/w). For proficiency, adequacy, and performance, PCR-QIA procedure is highly sensitive, a time-saver, electronically documented, mutagenic-reagent free, of little manual errors, accurate in measuring PCR fragments length, and quantitative data supplier. In conclusion, it can be suggested that optimized PCR-QAI is considered as a rapid and sensitive method for routine pork detection and quantification in raw or processed meat.
根据清真食品标签实施非常严格的程序，检测和量化“清真”肉类制品中的猪肉掺假是现代食品检验领域的一个关键问题。本研究旨在检测和定量生的和熟的加工香肠中的猪肉掺假。这是通过应用优化的种特异性 PCR 程序，然后进行 QIAxcel 毛细管电泳系统。制造实验设计为将猪肉与牛肉以 0.01 至 10%的替代水平混合，除了牛肉和猪肉香肠作为阴性和阳性对照。随后，将香肠分为生香肠和熟香肠，然后进行 DNA 提取。结果表明，通过猪特异性引物对线粒体 D 环和细胞色素 b (cytb)基因进行 PCR 扩增，分别在香肠中产生 185 和 117 bp 的猪肉特异性 DNA 片段。当 PCR 应用于牛肉香肠 DNA 时，未检测到 DNA 片段，证实了引物的特异性。作为内部对照，从猪肉和牛肉 DNA 模板中扩增出真核 18S 核糖体 RNA (rRNA)基因的 141-bp DNA 片段。尽管 PCR 后无论是 QIAxcel 还是琼脂糖技术都能有效地对目标 DNA 片段进行区分，即使是低至 0.01%（猪肉/肉：w/w）。为了胜任、充分性、性能，PCR-QIA 程序具有高度的敏感性、节省时间、电子记录、无诱变剂、手动错误少、准确测量 PCR 片段长度和提供定量数据的特点。总之，可以认为优化的 PCR-QAI 是一种快速、敏感的方法，可用于常规生肉或加工肉中的猪肉检测和定量。
