Full
Outline
To develop of a duplex real-time fluorescence polymerase chain reaction (PCR)-based method for the authentication of yak milk and yak-cattle milk.
Using species-specific nuclear chromosome genes of yak and cattle as targets, real-time PCR primers and probes were designed, and the duplex real-time fluorescence PCR system was optimized to achieve simultaneous detection of yak- and cattle-derived components in yak milk and yak-cattle milk.
The optimized duplex real-time fluorescence PCR system produced standard curves for yak milk (Y=–3.4015X+30.842, r2=0.9953) and cattle-yak milk (Y=–3.333X+30.035,r2=0.9984) showing excellent linearity, with amplification efficiencies of 96.78% and 99.54%, respectively. No cross-reactivity was observed with milk components from cattle, sheep, donkey, camel or other species, indicating strong specificity. In adulteration testing, the method successfully detected as low as 10% yak-cattle milk mixed into yak milk. Additionally, testing of 17 real samples confirmed the method’s high accuracy.
A duplex real-time fluorescence PCR method based on species-specific nuclear gene sequences is successfully developed in this study. This method enables accurate detection of yak-derived and yak-cattle-derived components in dairy products, providing a reliable approach for the authenticity identification of yak milk.
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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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