A multiplex PCR method for detection of five animal species in processed meat products using novel species-specific nuclear DNA sequences

Wang, Wenjun; Wang, Xiaokang; Zhang, Qingde; Liu, Zuhong; Zhou, Xiang; Liu, Bang

doi:10.1007/s00217-020-03494-z

Cited by 14 publications

(10 citation statements)

References 31 publications

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“…Nowadays, PCR-based techniques are the effective methods for species authentication. Real-time PCR techniques and microchip electrophoresis-dependent multiplex PCR methods require special infrastructures [ 11 , 29 , 30 ]; multiplex PCR assays through simple agarose gel analysis minimizes the cost drastically on a large scale and can be easily carried out to verify the identity of ingredients in foodstuffs [ 6 , 31 , 32 ]. As seen in Table 3 , much is known about multiplex PCR assays that simultaneously verify two to six meat ingredients in one reaction.…”

Section: Discussionmentioning

confidence: 99%

“…However, meat adulteration such as unlisted, mislabeled or fraudulent ingredients has frequently been reported around the world and has become a severe global issue [ 4 , 5 ]. Although some laws have been enacted for ensuring the quality and safety of meat products, adulteration is still widespread due to the purpose of economic pursuit [ 1 , 6 ]. Poultry meat (chicken, duck and goose), especially, is frequently adulterated to red meat due to their low cost of production in Chinese markets [ 1 , 7 ].…”

Section: Introductionmentioning

confidence: 99%

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A Simple and Reliable Single Tube Septuple PCR Assay for Simultaneous Identification of Seven Meat Species

Cai

Zhou

Liu

et al. 2021

Foods

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Multiplex PCR methods have been frequently used for authentication of meat product adulteration. Through screening of new species-specific primers designed based on the mitochondrial DNA sequences, a septuple PCR method is ultimately developed and optimized to simultaneously detect seven species including turkey (110 bp), goose (194 bp), pig (254 bp), sheep (329 bp), beef (473 bp), chicken (612 bp) and duck (718 bp) in one reaction. The proposed method has been validated to be specific, sensitive, robust and inexpensive. Taken together, the developed septuple PCR assay is reliable and efficient, not only to authenticate animal species in commercial meat products, but also easily feasible in a general laboratory without special infrastructures.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A Simple and Reliable Single Tube Septuple PCR Assay for Simultaneous Identification of Seven Meat Species

Cai

Zhou

Liu

et al. 2021

Foods

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“…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.…”

Section: Discussionmentioning

confidence: 99%

“…This study shows that PCR-HRM would be particularly useful in investigating admixture of vertebrate species in commercial processed meat products such as sausages, kebabs, meatballs and hams, which are frequently adulterated with multiple undeclared meats during processing (Wang et al, 2020). In performing such analysis, our findings underpin the need to employ more than one marker to ensure accurate species identification in meat admixtures.…”

Section: Discussionmentioning

confidence: 99%

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

Njaramba

Wambua

Mukiama

et al. 2021

Preprint

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Food fraud in several value chains including meat, fish, and vegetables has gained global interest in recent years. In the meat value chain, substitution of high commercial-value meats with similar cheaper or undesirable species is a common form of food fraud that raises ethical, religious, and dietary concerns. The presence of undeclared species could also pose public health risks caused by allergic reactions and the transmission of food-borne or zoonotic pathogens. Measures to monitor meat substitution are being put in place in many developed countries. However, information about similar efforts in sub-Saharan Africa is sparse. In this study, we used PCR coupled with high-resolution melting (PCR-HRM) analysis targeting the three mitochondrial genes, cytochrome oxidase 1 (CO1), cytochrome b (cyt b), and 16S rRNA, to detect species substitution in meat sold to consumers in Nairobi, Kenya’s capital city. Out of 107 meat samples from seven common livestock animals (cattle, goat, sheep, pig, chicken, rabbit, and camel), 11 (10.3%) had been substituted. Of 61 samples sold as beef, two were goat and one was camel. Of 30 samples sold as goat meat, four were mutton (sheep) and three were beef. One of nine samples purchased as pork was beef. Our results indicate that PCR-HRM analysis is a cost and time effective technique that can be employed to detect species substitution. The combined use of the three markers produced PCR-HRM profiles that successfully allowed the distinction of species. We demonstrate its utility not only in analysis of raw meat samples, but also of cooked, dried, and rotten samples, meat mixtures, and with the use of different DNA extraction protocols. We propose that this approach has broad applications in authentication of meat products and protection of consumers from food fraud in the meat industry in low- and middle-income countries such as Kenya, as well as in the developed world.

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“…The adulteration of meat with products from multiple species is increasingly being reported [ 17 , 45 , 46 , 47 , 48 ], thereby raising the demand for affordable and faster techniques for their detection. Many studies describing multi-species analyses of vertebrates in meat have utilized multiplex PCR [ 45 , 46 , 47 , 49 ]. While useful, multiplex PCR requires the use of expensive probes and post-PCR procedures, such as gel electrophoresis for the size separation of amplicons and/or DNA sequencing, thereby increasing analysis time, cost, and the risk of cross-contamination.…”

Section: Discussionmentioning

confidence: 99%

Detection of Species Substitution in the Meat Value Chain by High-Resolution Melting Analysis of Mitochondrial PCR Products

Njaramba

Wambua

Mukiama

et al. 2021

Foods

View full text Add to dashboard Cite

Substituting high commercial-value meats with similar cheaper or undesirable species is a common form of food fraud that raises ethical, religious, and dietary concerns. Measures to monitor meat substitution are being put in place in many developed countries. However, information about similar efforts in sub-Saharan Africa is sparse. We used PCR coupled with high-resolution melting (PCR-HRM) analysis targeting three mitochondrial genes—cytochrome oxidase 1 (CO1), cytochrome b (cyt b), and 16S rRNA—to detect species substitution in meat sold to consumers in Nairobi, Kenya. Out of 107 meat samples representing seven livestock animals, 11 (10.3%) had been substituted, with the highest rate being observed in samples sold as goat. Our results indicate that PCR-HRM analysis is a cost- and time-effective technique that can be employed to detect species substitution. The combined use of the three mitochondrial markers produced PCR-HRM profiles that successfully allowed for the consistent distinction of species in the analysis of raw, cooked, dried, and rotten meat samples, as well as of meat admixtures. We propose that this approach has broad applications in the protection of consumers against food fraud in the meat industry in low- and middle-income countries such as Kenya, as well as in developed countries.

show abstract

A multiplex PCR method for detection of five animal species in processed meat products using novel species-specific nuclear DNA sequences

Cited by 14 publications

References 31 publications

A Simple and Reliable Single Tube Septuple PCR Assay for Simultaneous Identification of Seven Meat Species

A Simple and Reliable Single Tube Septuple PCR Assay for Simultaneous Identification of Seven Meat Species

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

Detection of Species Substitution in the Meat Value Chain by High-Resolution Melting Analysis of Mitochondrial PCR Products

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