Full
Outline
To establish a novel method for rapid on-site detection of the tetracycline resistance gene TetA by cross priming amplification (CPA) technology.
A total of 5 specific primers targeting conserved regions of the TetA gene were designed. Amplification was performed under isothermal conditions (optimized at 51–60 ℃), with the entire detection process completed within 45–60 minutes using only basic heating equipment such as a water bath or metal bath. To validate its performance, the method was systematically evaluated for sensitivity, specificity and interference resistance.
The limit of detection for purified TetA gene templates was as low as 30.0 fg/μL, indicating high sensitivity. Furthermore, no nonspecific amplification was observed in comparative tests against other tetracycline resistance genes (e.g., tetB, tetM) and common clinical pathogenic genes, confirming excellent selectivity. Additionally, simulated sample testing yielded stable and reliable results.
A simple, rapid, low-cost and instrument-free CPA-based method for TetA gene detection is successfully developed. With its high sensitivity and specificity, this method can effectively meet the needs for rapid on-site screening of tetracycline resistance genes in primary healthcare institutions and farming environments. It provides a robust technical tool for real-time monitoring and controlling the spread of antibiotic resistance genes, demonstrating significant practical application value.
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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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