Mycobacterium tuberculosis rpoB gene mutation detection method and kit based on CRISPR-Cas12a technology
By designing a specific crRNA binding with Cas12a enzyme and fluorescent probe, the problems of complexity, time consumption and insufficient sensitivity of traditional detection methods are solved, enabling rapid, sensitive and specific detection of Mycobacterium tuberculosis rpoB gene mutations, which is suitable for detecting low-abundance mutations in mixed samples.
Patent Information
- Application Number
- CN202511150785.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-18
- Publication Date
- 2025-11-21
AI Technical Summary
Traditional detection methods for Mycobacterium tuberculosis rpoB gene mutations are complex, time-consuming, and lack sensitivity in mixed samples. CRISPR-Cas12a technology has limitations in crRNA design optimization and PAM sequence restrictions in the detection of drug resistance mutations in Mycobacterium tuberculosis, making it difficult to achieve high specificity and high sensitivity.
The design incorporates specific crRNAs that are tightly paired with rpoB gene mutation sites, combined with Cas12a enzyme and fluorescent probes. Rapid and sensitive mutation detection is achieved through PCR amplification and a CRISPR-Cas12a detection system, making it particularly suitable for detecting low-abundance mutations in mixed samples.
It enables rapid, sensitive, and specific detection of drug-resistant mutations in Mycobacterium tuberculosis, with a detection time of ≤2 hours and a sensitivity of up to 0.1%. The detection results are highly consistent with Sanger sequencing, making it suitable for point-of-care clinical testing.