Simple and fast multiplex PCR method for detection of species origin in meat products

Abstract

Identification of animal species is one of the major concerns in food regulatory control and quality assurance system. Different approaches have been used for species identification in animal origin of feedstuff. This study aimed to develop a multiplex PCR approach to detect the origin of meat and meat products. Specific primers were designed based on the conserved region of mitochondrial Cytochrome C Oxidase subunit I () gene. This method could successfully distinguish the origin of the pig, camel, sheep, donkey, goat, cow, and chicken in one single reaction. Since PCR products derived from each species represent unique molecular weight, the amplified products could be identified by electrophoresis and analyzed based on their size. Due to the synchronized amplification of segments within a single PCR reaction, multiplex PCR is considered to be a simple, fast, and inexpensive technique that can be applied for identification of meat products in food industries. Nowadays, this technique has been considered as a practical method to identify the species origin, which could further applied for animal feedstuffs identification.

The Quality of DNA Isolated from Processed Food and Feed via Different Extraction Procedures

“…Thus, DNA-based analytical methods represent a crucial approach for species identification in raw and highly processed meat products due to their high thermal stability and their very low detection limits. PCR and real-time PCR proved to be fast, reliable, and sensitive methods for species identification in foods and feeds with complex compositions [7,8,9,10,11,12,13,14]. Sequences of mitochondrial [15,16,17] and genomic DNA [18] were reported to be targeted as markers.…”

Species substitution in the meat value chain by high-resolution melt analysis of mitochondrial PCR products

“…Nevertheless, our previous studies show that PCR-HRM analysis provides a degree discovery of novel/uncharacterized species, which can be identified by DNA sequencing (Ouso et al, 2020). Furthermore, the mitochondrial markers CO1 and cyt b , are often used singly in species identification (Di Pinto et al, 2015; Lopez-Oceja et al, 2017; Izadpanah et al, 2018), but our results indicate the need to use multiple markers in tandem for accurate species identification. Finally, mitochondrial markers which are commonly used for DNA barcoding of species, are not appropriate for quantification of species adulteration in meat mixtures because there are major differences in gene copies of mtDNA markers in different species (Ballin et al, 2009; Cai et al, 2017).…”

“…Adulteration of meat with products from multiple species is increasingly being reported (Ali et al, 2015; Di Pinto et al, 2015; Izadpanah et al, 2018; Kitpipit et al, 2014; O’Mahony, 2013), raising the demand for affordable and faster techniques for their detection. Many studies describing multi-species analysis of vertebrates in meat have utilized multiplex PCR (Ali et al, 2015; Balakrishna et al, 2019; Izadpanah et al, 2018; Kitpipit et al, 2014; Li et al, 2019; Liu et al, 2019; Qin et al, 2019; Wang et al, 2020). While useful, multiplex PCR requires use of expensive probes and post-PCR procedures such as agarose gel electrophoresis for size separation of amplicons and/or DNA sequencing, thereby increasing analysis time, cost, and risk of cross-contamination.…”

Application of Histological and Physico-Chemical Analyses for Evaluation of Meat Product — Cachir

“…The identification of other ingredients used in the production of meat products is also important for the evaluation of the quality of the final product (Pospiech et al, 2009). Therefore, various molecular biology (Doosti et al, 2011;Izadpanah et al, 2017, Di Pinto et al, 2015 and histological methods (Sadeghinezhad et al, 2015;Abdel Hafeez et al al., 2016) have been developed to detect plant additives in meat products. The most important additives that can be studied with the use of histological techniques are plant additives such as plant tissues and starches.…”