Full
Outline
To develop a meat component detection method based on handheld quantitative real-time polymerase chain reaction (PCR) for rapid on-site identification of meat sources.
This study focused on the pork, chicken and duck-derived components, explored the influence of the single factors of meat sample pretreatment tools, lysate composition, lysate concentration and lysate addition amount on the extracted DNA, and constructed a rapid pretreatment method for meat samples; screened specific primers and probes, used the handheld quantitative teal-time PCR instrument to verify specificity, sensitivity and limit of detection, and drew amplification standard curves; and detected the real single and mixed meat samples.
The established rapid pretreatment method for meat samples could be completed within 5 min, the DNA extraction took 10 min, and the extracted DNA could meet the requirements of subsequent experiments. The detection methods exhibited high specificity for pork,chicken and duck samples. The limits of detection of pork, chicken and duck-derived DNA components were 0.010,0.001 and 0.001 ng/µL. The standard curve showed a sound linear relationship. The amplification standard curve of pig DNA was: Y=–3.956X+38.803, r2=0.9956. The amplification standard curve of chicken DNA was: Y=–3.499X+40.491,r2=0.9909. The amplification standard curve of duck DNA was: Y=–3.961X+39.192, r2=0.9962. The method was successfully applied to detect 28 meat samples with a 100% success rate.
These results indicate that the rapid meat sample pretreatment method established in this experiment can be used for the rapid and accurate detection of pork, chicken and duck meat on a handheld quantitative real-time PCR all-in-one machine. This approach provides robust technical support for monitoring and controlling meat adulteration at the retail or market level.
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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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