PDF
Full
Outline
Red fire ant, Solenopsis invicta, a serious invasive pest native to South America, has been widely distributed in China, posing significant threats to agriculture, natural ecosystems, and public health. Effective entry-exit quarantine measures are essential to prevent the invasion and spread of S. invicta, which requires rapid and accurate identification of the pest. Existing detection methods, including morphological identification, DNA barcoding, immunoassay, and loop-mediated isothermal amplification (LAMP) have limitations in simplicity, detection speed and sensitivity. Recombinase polymerase amplification (RPA) is an isothermal amplification technique characterized by its simplicity, high sensitivity, and efficient amplification. To enable rapid and accurate on-site detection of S. invicta, this study developed a visual detection method based on RPA technology. Mitochondrial genome sequences of S. invicta and closely related or morphologically similar species were analyzed using data from the NCBI database. The mitochondrial NADH2 dehydrogenase gene was selected as the target for amplification, and specific RPA primers were designed. Experimental results confirmed that the primers demonstrated high specificity for S. invicta. Sensitivity testing using agarose gel electrophoresis revealed that the RPA reaction could detect as low as 1.0×10-4 ng/μL, which is 103 times more sensitive than conventional PCR. A visual detection method was further developed by incorporating nucleic acid dyes into the reaction, achieving a sensitivity of approximately 1.0×10-1 ng/μL. Additionally, a rapid DNA extraction method was explored. This method involved grinding ant samples in sterile water followed by heating at 100 ℃ in a water bath for one minute, which successfully yielding crude DNA. The developed method enables gene amplification of S. invicta at 37 ℃, with detection results assessed via fluorescence reactions, completing the entire process within 30 minutes. This approach provides a convenient, rapid, accurate, and reliable technical method for detecting S. invicta, offering valuable technical support for its quarantine, prevention and control.
PDF
107
54
| 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 |
关闭全屏
No. 86 Xueyuan South Road, Haidian District, Beijing
010-62199257
qkjq@cast.org.cn
Copyright © 2025 China Association for Science and Technology. All rights reserved. For all open access content, the relevant licensing terms apply.
Sponsored by the Office of the Leading Group for Cybersecurity and Informatization of CAST, and supported by Science and Technology Review Publishing House
