Full
Outline
To establish a method for determination of single-copy nuclear gene targets of animal-derived components in food by real-time fluorescence quantitative polymerase chain reaction (qPCR).
qPCR primers and Taqman probes were designed to target the specific nuclear genes of common animal-derived components in food to verify and confirm the specificity and sensitivity of the method. The applicability and stability of simulated and actual commercially available animal-derived food detection methods were adopted, and the qPCR detection values were compared and analyzed with those of mitochondrial cytoplasmic gene target detection.
The qPCR primer probe for single-copy nuclear gene targets had good specificity, with a minimum limit of detection of 0.1 ng/μL DNA mass concentration, nuclear genes could accurately identify the minimum adulteration component content of 0.1% (mass ratio). The cycle threshold (Ct) value of the nuclear genes amplification reactions were all greater than that of the cytoplasmic target gene, and the amplification reactions of cytoplasmic genes could recognize a lower adulteration content of 0.01% (mass ratio). In the testing of simulated and actual commercially available samples, the nuclear gene qPCR reaction could accurately and effectively detect animal-derived samples mixed with different source components, and it was not easy to detect chicken essence ingredients and other source contamination components from the production line.
The research results of this paper show that the qPCR amplification system established based on single-copy nuclear gene targets has strong specificity and high sensitivity for the detection of common animal-derived components in food. Compared with the amplification results of mitochondria genes, the nuclear genes’ detection method is less affected by the substitution of ingredients and the cross-contamination on the production line, and can quickly and accurately detect animal-derived component in food, which make it conducive to market supervision and risk assessment.
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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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