Development and Evaluation of Elisa Kits Based on Proteomics Technology for Detection and Semi-Quantification of Meat Species Adulteration

M. M. Ahmed, Walid Nazmy, A. Noureen
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Abstract

Purpose: This study aimed to develop ELISA kits for the detection of meat species adulteration in raw and processed meat and evaluate its specificity and sensitivity. Method: We identify the unique peptide markers based on the proteomics approach. Subsequently, we prepared specific antisera for beef, pork, horse, sheep, chicken and developed ELISA kits. Results: Evaluation of the developed kits in testing crooked reference sample mixtures revealed that the developed  ELISA kits showed 100% specificity with no cross-reactivity detected for the non-target species. In terms of sensitivity, the developed ELISA kits were able to consistently detect pork in the sample mixtures and horses at levels down to 5% w/w. Although sheep and chicken were detected at levels as low as 1% w/w. The beef-specific ELISA test showed greater sensitivity.   Conclusions: It is concluded that the developed ELISA kits are a promising tool but further studies are still required for validation of the developed kits and comparing its results with the commercially used kits before entry into the production phase.
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基于蛋白质组学技术的肉品掺假半定量检测试剂盒的研制与评价
目的:建立生肉和加工肉制品中肉类种类掺假的酶联免疫吸附测定试剂盒,并对其特异性和敏感性进行评价。方法:采用蛋白质组学方法鉴定独特的肽标记。随后,我们制备了牛肉、猪肉、马、羊、鸡的特异性抗血清,并开发了ELISA试剂盒。结果:开发的试剂盒对弯曲参比样品混合物的检测结果表明,开发的ELISA试剂盒对非靶种的特异性为100%,无交叉反应。在灵敏度方面,开发的ELISA试剂盒能够始终如一地检测样品混合物中的猪肉和马,含量低至5% w/w。尽管绵羊和鸡的含量低至1% w/w。牛肉特异性ELISA检测显示出更高的敏感性。结论:开发的ELISA试剂盒是一种很有前途的工具,但在进入生产阶段之前,仍需要进一步的研究来验证开发的试剂盒,并将其结果与商业使用的试剂盒进行比较。
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