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Outline
Objective: To establish a multiplex real-time PCR assay capable of simultaneously and rapidly identifying bovine, chicken, duck and porcine components in beef products and thus to counteract economically motivated adulteration with cheaper meats. Methods: Species-specific primers and TaqMan probes targeting the mitochondrial cytochrome b (Cytb) gene were designed and screened for cattle, chicken, duck and pig. A single-tube reaction system was constructed and systematically validated for sensitivity, specificity, linearity and adulteration simulation. Results: The assay exclusively amplified DNA from cattle, chicken, duck and pig, with no cross-reactivity against 6 non-target species. The method simultaneously detected four animal-derived DNA with 10−4 ng/μL sensitivity and a 0.01% (w/w) detection limit. Screening of 100 retail beef samples revealed 24 adulterated products, predominantly containing duck meat and/or pork. All results were fully concordant with those obtained by the national standard method. Conclusion: The developed multiplex real-time PCR protocol is rapid, cost-effective and high-throughput, significantly enhancing the capability to detect meat adulteration and providing robust technical support for food-safety regulation and consumer protection.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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