논문
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In this project, we established a rapid room-temperature pretreatment method for sample extraction and improved Cas12a-based technique to develop an extraction-free one-pot pathogen detection method. This novel approach, termed EXORCA (EXtraction-free One-pot RPA-CRISPR/Cas12a assay), significantly improves the convenience of existing diagnostic tools and addresses the current limitations currently associated with diagnosing K. pneumoniae. EXORCA allows for detecting K. pneumoniae within approximately 30 min using unextracted samples. In addition, it incorporates blue light for visual detection, enhancing its applicability in POCT. Taking full advantage of the specificity of Cas12a, EXORCA can distinguish K. pneumonia from other strains with 100% accuracy. Clinical sample tests have demonstrated that the EXORCA assay aligns perfectly with qPCR results, showing 100% (20/20) consistency. These results indicate that the EXORCA assay holds significant promise as a POCT tool for the rapid and accurate detection of K. pneumonia
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In this project, we established a rapid room-temperature pretreatment method for sample extraction and improved Cas12a-based technique to develop an extraction-free one-pot pathogen detection method. This novel approach, termed EXORCA (EXtraction-free One-pot RPA-CRISPR/Cas12a assay), significantly improves the convenience of existing diagnostic tools and addresses the current limitations currently associated with diagnosing K. pneumoniae. EXORCA allows for detecting K. pneumoniae within approximately 30 min using unextracted samples. In addition, it incorporates blue light for visual detection, enhancing its applicability in POCT. Taking full advantage of the specificity of Cas12a, EXORCA can distinguish K. pneumonia from other strains with 100% accuracy. Clinical sample tests have demonstrated that the EXORCA assay aligns perfectly with qPCR results, showing 100% (20/20) consistency. These results indicate that the EXORCA assay holds significant promise as a POCT tool for the rapid and accurate detection of K. pneumonia