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The global proliferation of carbapenemase-producing bacteria (CPB) has garnered significant attention worldwide. Early diagnosis of CPB and accurate identification of carbapenemases are crucial for preventing the spread of CPB and ensuring targeted antibiotic therapy. Therefore, efficient and accurate identification of carbapenemases is paramount in clinically treating diseases associated with CPB. In this study, 58 CPB strains were collected and detected using the DNA endonuclease-targeted CRISPR trans reporter (DETECTR) method, a rapid detection platform based on CRISPR-Cas12a gene editing and isothermal amplification. Additionally, four conventional methods (the APB/EDTA method, PCR, NG-test Carba 5, and GeneXpert Carba-R) were employed and compared against whole genome sequencing (WGS) results, considered the gold standard, to evaluate their efficacy in detecting carbapenemases. Detection by the APB/EDTA method revealed that 29 strains were positive for Class A serine endopeptidases, while 29 strains were positive for Class B metalloenzymes. The classification of these zymotypes was consistent with the sequencing result. All target carbapenemases for KPC were identified with 100% sensitivity using NG-test Carba 5, PCR, DETECTR, and GeneXpert Carba-R. In the case of NDM, both Xpert Carba-R and DETECTR showed a sensitivity of 100%. In contrast, NG-test Carba 5 and PCR had a slightly lower sensitivity of 96.7%, each missing one target carbapenemase. n this study, the APB/EDTA method is capable of identifying the zymotype classification but not the specific resistant genes, while Xpert Carba-R and DETECTR are able to detect all target carbapenemases.

The recent major worldwide outbreak of monkeypox virus (MPXV) has highlighted the urgent need for accurate MPXV detection methods. Although quantitative PCR (qPCR) technique is currently the gold standard for MPXV diagnosis, the high costs associated with the technique and the need for complex instrumentation, limits its application in resource‐poor settings. CRISPR technology has developed rapidly in recent years and provides an effective tool for point‐of‐care testing pathogen identification. Here, we exploited the cleavage properties of the Cas12a enzyme and Cas13a enzyme, to detect the MPXV specific genes, F3L gene and B6R gene, respectively. We developed two detection protocols: a 2‐step method in which the CRISPR Dual System reaction and the multiplex recombinase polymerase amplification reaction were carried out in separate tubes and a single‐tube method in which both reactions were carried out in one tube. Evaluation of the two methods showed that our protocol can detect the MPXV genome down to 10° copies/μL with good specificity and no cross‐reactivity with other poxviruses pseudoviruses, and bacteria. Mock positive samples were used to assess clinical applicability, with the results showing satisfactory concordance with the qPCR method for parallel testing. In conclusion, our study provides a reliable molecular diagnostic strategy for detection of MPXV. [ABSTRACT FROM AUTHOR]