Nur Fadhilah Khairil Mokhtar, Ying Quah Shun, Raja Mohd Hafidz Raja Nhari, Nurhidayatul Asma Mohamad, Nur Maisarah Shahidan, Irwan Hanish Warsanah, Amalia Mohd Hashim
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引用次数: 0
Abstract
The inclusion of ingredients derived from pigs in highly processed consumer products poses a significant challenge for DNA-targeted analytical enforcement, which could be overcome by using digital PCR. However, most species detection methods use digital PCR to target single-copy nuclear genes, which limits their sensitivity. In this work, we examined the performance of a nanoplate-based digital PCR method that targets multi-copy nuclear (MPRE42) and mitochondrial (Cytb) genes. Poor separation of positive and negative partitions, as well as a 'rain effect' were obtained in the porcine-specific MPRE42 assay. Among the optimization strategies examined, the inclusion of restriction enzymes slightly improved the separation of positive and negative partitions, but a more extensive 'rain effect' was observed. The high copy number of the MPRE42 amplicon is hypothesized to contribute to the saturation of the positive signal. In contrast, the porcine-specific Cytb assay achieved perfect separation of positive and negative partitions with no 'rain effect'. This assay can detect as little as 0.4 pg of pork DNA, with a sensitivity of 0.05% (w/w) in a pork-chicken mixture, proving its applicability for detecting pork in meat and meat-based products. For the MPRE42 assay, potential applications in highly degraded products such as gelatin and lard are anticipated.
期刊介绍:
Food Additives & Contaminants: Part A publishes original research papers and critical reviews covering analytical methodology, occurrence, persistence, safety evaluation, detoxification and regulatory control of natural and man-made additives and contaminants in the food and animal feed chain. Papers are published in the areas of food additives including flavourings, pesticide and veterinary drug residues, environmental contaminants, plant toxins, mycotoxins, marine biotoxins, trace elements, migration from food packaging, food process contaminants, adulteration, authenticity and allergenicity of foods. Papers are published on animal feed where residues and contaminants can give rise to food safety concerns. Contributions cover chemistry, biochemistry and bioavailability of these substances, factors affecting levels during production, processing, packaging and storage; the development of novel foods and processes; exposure and risk assessment.