Full
Outline
To establish n method for simultaneously identifying multiple meat species within a single reaction system by efficient multiplex polymerase chain reaction (PCR) technology.
Based on species-specific variations in animal mitochondrial gene sequences, primers were designed and screened using multiple bioinformatics tools, and an octuple PCR assay was developed for the simultaneous detection of 8 kinds of meat species, namely mutton, chicken, rabbit, pigeon, goose, turkey, ostrich and dog. The specificity of the amplification products was confirmed through reaction condition optimization and DNA sequencing. The sensitivity and applicability of the method were further evaluated by multi-species DNA mixtures and adulterated meat model samples.
The octuple PCR assay enabled stable and simultaneous amplification of DNA from all 8 kinds of target species in a single reaction tube, with no nonspecific amplification observed. Sequencing results were consistent with the corresponding target species. Sensitivity analysis showed that, in both raw and heat-processed meat samples, the limits of detection for the target species ranged from 0.01 to 0.05 ng of genomic DNA. In adulterated samples, target meat components were detectable at levels as low as 0.10%.
The established octuple PCR method can be successfully applied to the authenticity assessment of commercially available meat products, allowing rapid and accurate identification of multiple meat species. This method demonstrates good practical applicability and provides effective technical support for food safety surveillance and market regulation.
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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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