Primer probe combination, kit and method for onsite detection of rana spinosa based on RPA (recombinase polymerase amplification) technology
By designing highly specific and sensitive RPA primer-probe combinations and nucleic acid test strips, combined with DNA isothermal rapid amplification technology, the problem of rapid detection of spotted tree frogs in the field has been solved, achieving simple, rapid, and accurate detection results.
CN122445808APending Publication Date: 2026-07-24SHANGHAI WILDLIFE & NATURE RESERVE RES CENT
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Patent Information
- Application Number
- CN202610775691.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-06-01
- Publication Date
- 2026-07-24
AI Technical Summary
Technical Problem
Existing technologies are insufficient for the rapid and accurate detection of the spotted tree frog in the wild. Traditional PCR methods are time-consuming and have high environmental requirements, making real-time monitoring impossible.
Method used
We designed a highly specific and sensitive RPA primer-probe composition, combined with nucleic acid detection test strips, and used DNA isothermal rapid amplification technology to achieve on-site detection of spotted rain frogs.
Benefits of technology
The test can produce results within 50 minutes at 38℃. It is easy to operate, highly sensitive and specific, and suitable for rapid monitoring of spotted rain frogs in the field.
✦ Generated by Eureka AI based on patent content.
Abstract
The invention discloses a primer probe combination, a kit and a method for onsite detection of rana spinosa on the basis of an RPA (recombinase polymerase amplification) technology. The primer probe composition provided by the invention consists of an upstream primer, a downstream primer and a probe, the upstream primer is as shown in SEQ ID NO: 1 or SEQ ID NO: 4; the downstream primer is as shown in SEQ ID NO: 2 or SEQ ID NO: 5; the probe is as shown in SEQ ID NO: 3 or SEQ ID NO: 6. The invention provides a method for detecting whether spot-free rain frogs exist in a water body environment. The method comprises the following steps: (1) taking a water sample from the water body environment and extracting DNA (Deoxyribose Nucleic Acid); (2) taking the DNA extracted in the step (1) as a template, and carrying out RPA amplification by adopting the primer probe composition; and (3) taking an amplification product in the step (2), detecting by adopting a nucleic acid detection test strip, and reading a result. The invention establishes the RPA-LFD detection method for the spotless frog, and the RPA-LFD detection method is suitable for field rapid monitoring.
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