Identification of eleven meat species in foodstuff by a hexaplex real-time PCR with melting curve analysis

Li, Jinchun; Li, Jiapeng; Liu, Ruixi; Wei, Yixuan; Wang, Shouwei

doi:10.1016/j.foodcont.2020.107599

Cited by 34 publications

(17 citation statements)

References 30 publications

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“…In addition, meat fraud risks food safety and even threatens public health because of metabolic disorders, allergies, or infectious diseases. As reported, forbidden ingredients such as fox and mouse are occasionally mixed into edible meat products ( 3 , 8 ). Nevertheless, allergic reactions can be triggered by some meat species especially for sensitized patients ( 9 ).…”

Section: Introductionmentioning

confidence: 99%

Molecular Authentication of Twelve Meat Species Through a Promising Two-Tube Hexaplex Polymerase Chain Reaction Technique

et al. 2022

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Frequent meat frauds have aroused significant social attention. The aim of this study is to construct a two-tube hexaplex polymerase chain reaction (PCR) method offering accurate molecular authentication of twelve meat species in actual adulteration event. Deoxyribonucleic acid (DNA) sequencing demonstrates that designed primers can specifically amplify target species from genomic DNA mixture of six species in each tube reaction, which showed 100% accuracy of horse (148 bp), pigeon (218 bp), camel (283 bp), rabbit (370 bp), ostrich (536 bp), and beef (610 bp) as well as turkey (124 bp), dog (149 bp), chicken (196 bp), duck (277 bp), cat (380 bp), and goose (468 bp). A species-specific primer pair produced the target band in the presence of target genomic DNA but not non-target species. Through multiplex PCR assays with serial concentration of the DNA mixture of six species in each PCR reaction, the detection limit (LOD) of the two-tube hexaplex PCR assay reached up to 0.05–0.1 ng. Using genomic DNA isolated from both boiled and microwave-cooked meat as templates, PCR amplification generated expected PCR products. These findings demonstrate that the proposed method is specific, sensitive and reproducible, and is adequate for food inspection. Most importantly, this method was successfully applied to detect meat frauds in commercial meat products. Therefore, this method is of great importance with a good application foreground.

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Section: Introductionmentioning

confidence: 99%

Molecular Authentication of Twelve Meat Species Through a Promising Two-Tube Hexaplex Polymerase Chain Reaction Technique

et al. 2022

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“…[1][2][3][4][5] Accurate diagnosis of disease-causing gene mutation is vital for clinical treatment, genetic counseling, and disease prevention strategies. Molecular techniques, including DNA sequencing, 6 probe hybridization, 7 capillary electrophoresis, 8 and melting curve analysis (MCA), 9 can identify more in-depth genetic information from the polymerase chain reaction (PCR) products. Among them, MCA is popular for single-tube detection of single-nucleotide polymorphism (SNP) 10,11 due to its simplicity, high sensitivity, and specificity.…”

Section: Introductionmentioning

confidence: 99%

One-shot high-resolution melting curve analysis for KRAS point-mutation discrimination on a digital microfluidics platform

Wan

Law

et al. 2022

Lab Chip

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“…Specifically, since the European horsemeat scandal in 2013, the events regarding ingredient substitution of expensive meat with low-cost ones for extra economic benefits have frequently occurred worldwide (3). The false information on the label will mislead the choices of consumers, resulting in serious religious issues and health problems (4,5). For example, meat products harboring pork ingredients are not permitted in Islamic countries (6,7).…”

Section: Introductionmentioning

confidence: 99%

A Heptaplex PCR Assay for Molecular Traceability of Species Origin With High Efficiency and Practicality in Both Raw and Heat Processing Meat Materials

Zhou

Zhong²,

Zhou³

et al. 2022

Front. Nutr.

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Frequent meat frauds have become a global issue because adulteration risks the food safety, breaches market rules, and even threatens public health. Multiplex PCR is considered to be a simple, fast, and inexpensive technique that can be applied for the identification of meat products in food industries. However, relatively less is known about a multiplex PCR method authenticating seven animal species simultaneously in one reaction due to technological challenge. Through screening new species-specific primers and optimizing PCR system, a heptaplex PCR method was established, which could simultaneously detect seven meat ingredients of camel (128 bp), pigeon (157 bp), chicken (220 bp), duck (272 bp), horse (314 bp), beef (434 bp), and pork (502 bp) in a single-tube reaction. DNA sequencing solidly validated that each set of primers specifically amplified target species from total DNA mixtures of seven meat species. The developed multiplex assay was stable and sensitive enough to detect 0.01–0.025 ng DNA from various meat treatments including raw, boiled, and autoclaved meat samples or target meat content of 0.1% total meat weight, suggesting the suitability of the heptaplex PCR technique for tracing target meats with high accuracy and precision. Most importantly, a market survey validated the availability of this multiplex PCR technique in real-world meat products with a good application foreground.

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Identification of eleven meat species in foodstuff by a hexaplex real-time PCR with melting curve analysis

Cited by 34 publications

References 30 publications

Molecular Authentication of Twelve Meat Species Through a Promising Two-Tube Hexaplex Polymerase Chain Reaction Technique

Molecular Authentication of Twelve Meat Species Through a Promising Two-Tube Hexaplex Polymerase Chain Reaction Technique

One-shot high-resolution melting curve analysis for KRAS point-mutation discrimination on a digital microfluidics platform

A Heptaplex PCR Assay for Molecular Traceability of Species Origin With High Efficiency and Practicality in Both Raw and Heat Processing Meat Materials

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