Kit for rapidly detecting mycoplasma synoviae based on ERA-CRISPR / Cas12a one-step method, application and method
By combining ERA isothermal amplification technology and CRISPR/Cas12a technology, the specific DNA target scpA was selected, and the one-pot method ERA-CRISPR/Cas12a technology was established, which solved the problems of cumbersome, time-consuming and low sensitivity of the existing detection methods, and achieved rapid detection of Mycoplasma of Chicken Synovial Sacculoplasm with high sensitivity and strong specificity.
CN119710048APending Publication Date: 2025-03-28SHANGHAI VETERINARY RESEARCH INSTITUTE CAAS (CHINESE ANIMAL HEALTH & EPIDEMIOLOGY CENTER SHANGHAI BRANCH)
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Patent Information
- Application Number
- CN202411980863.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-31
- Publication Date
- 2025-03-28
AI Technical Summary
Technical Problem
The existing methods of detecting Mycoplasma of chicken synovial fluid are cumbersome, time-consuming, and have low sensitivity, making it difficult to meet the needs of rapid on-site testing.
Method used
Combined with ERA isothermal amplification technology and CRISPR/Cas12a technology, through the selection of specific DNA target scpA, a one-pot ERA-CRISPR/Cas12a technology was established to quickly and accurately detect Mycoplasma of the chicken synovial fluid.
Benefits of technology
It realizes high sensitivity, strong specificity and rapid detection, and can detect mycoplasma of chicken synovial fluid as low as 3.81pg/μL. It is suitable for on-site rapid inspection and does not require expensive instruments and equipment.
✦ Generated by Eureka AI based on patent content.
Abstract
The invention discloses a kit, application and method for rapidly detecting mycoplasma synoviae based on an ERA-CRISPR / Cas12a one-step method, according to the visual detection kit, on the basis of a new nucleic acid detection target scpA of the mycoplasma synoviae, a sample to be detected is split by a splitting buffer to release genome DNA, then ERA-CRISPR / Cas12a one-pot reaction is carried out in a constant-temperature water bath kettle or a metal bath, and the sample to be detected is subjected to one-pot reaction, so that the ERA-CRISPR / Cas12a one-pot reaction is carried out to rapidly detect the mycoplasma synoviae through the ERA-CRISPR / Cas12a one-pot reaction. Under the mediation of a crRNA guide sequence, after Lb Cas12a enzyme is combined with crRNA and target DNA to form a ternary complex, the trans-shear activity of cas12a is activated, an ssDNA fluorescent probe in a reaction system can be arbitrarily cut to release a fluorescent group on the probe, and an ultraviolet lamp is used for detecting a fluorescent signal or a colloidal gold test strip is used for detecting whether FAM and Biotin groups are cut off for result judgment. According to the ERA-CRISPR / cas12a one-pot method technology for rapidly detecting the mycoplasma synoviae, the DNA target of the mycoplasma synoviae is recognized and cut through ERA amplification and CRISPR / cas12a specificity, the specificity is high, the detection sensitivity is as low as 3.81 pg / mu L, and the ERA-CRISPR / cas12a one-pot method technology can be used for rapidly detecting the mycoplasma synoviae in clinical samples on site.
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