A composition, method, and application for simultaneous detection of Staphylococcus aureus and Salmonella based on CRISPR-Cas12a / Cas12b.

By using RPA-CRISPR-Cas12a/Cas12b technology, combined with carefully designed crRNA and sgRNA, and optimizing the reaction system, the problems of cross-interference and stability in the detection of Staphylococcus aureus and Salmonella have been solved, achieving rapid and accurate dual detection, which is suitable for food safety testing.

CN122081529BActive Publication Date: 2026-07-17NANJING FOOD & DRUG SUPERVISION & INSPECTION INST
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
NANJING FOOD & DRUG SUPERVISION & INSPECTION INST
Filing Date
2026-04-22
Publication Date
2026-07-17

AI Technical Summary

Technical Problem

Existing technologies suffer from cross-interference and low stability when detecting Staphylococcus aureus and Salmonella. Furthermore, traditional methods are highly dependent on equipment and involve cumbersome procedures, making it difficult to achieve rapid and accurate multiplex detection.

Method used

Using RPA-CRISPR-Cas12a/Cas12b technology, and through carefully designed highly specific crRNA and sgRNA combined with thiomodified probes, we can achieve simultaneous detection of Staphylococcus aureus and Salmonella. We have optimized the reaction system and parameters to improve the specificity and sensitivity of the method.

Benefits of technology

It enables rapid and accurate detection of Staphylococcus aureus and Salmonella within 1 hour, with good specificity and high sensitivity. It is suitable for food safety testing in different scenarios and avoids equipment dependence and cumbersome operation.

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Abstract

This invention discloses a composition, method, and application for the simultaneous detection of Staphylococcus aureus and Salmonella based on CRISPR-Cas12a / Cas12b, belonging to the field of food safety testing technology. Based on the characteristics of RPA isothermal amplification and the preference for different Cas12 enzyme cleavage sites, this invention establishes a dual RPA-CRISPR-Cas12a / Cas12b detection method for Staphylococcus aureus and Salmonella through primer screening and optimization of parameters such as crRNA, reaction time, and temperature. This effectively solves the problem of simultaneous detection of multiple DNA targets in the Cas12 system. This invention exhibits good specificity, high sensitivity, and simple result interpretation, achieving a detection limit of 10 for both pure cultures and artificially contaminated samples. 0 With a CFU / mL level, detection can be completed within 1 hour, providing a new technical approach for rapid screening of foodborne pathogens.
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