Detection of KPC-Producing Carbapenem-Resistant Klebsiella pneumoniae Based on CRISPR Cas12a
Year: 2025
Type: article
Abstract: To develop a detection system for Klebsiella pneumoniae carbapenemase (KPC) and provide a reference for clinical prevention and control of nosocomial infections caused by multidrug-resistant K. pneumoniae.The KPC resistance gene was amplified by PCR.Guided by crRNA, Cas12a specifically identified the resistance gene and activated its trans-cleavage activity.In the detection system, a fluorescence probe was cleaved by activated Cas12a, and the fluorescence signal was measured using a microplate reader.Under optimized conditions, the fluorescence signal appeared within 12 min, peaked at 40 min and completed detection within 60 min.sensitivity: 91.2%, specificity: 84.1%, detection limit: 0.01 ng/μl.The samples were examined by fluorescence-CRISPR Cas12a and PCR.The coincidence rate was 85.9%, Kappa value was 0.8.The ROC curve analysis revealed an AUC of 0.916, with an optimal cutoff value of 1.55, sensitivity of 91.2%, and specificity of 84.1%.The CRISPR Cas12a detection of carbapenem-resistant K. pneumoniae (CRKP) demonstrates high sensitivity, specificity, and broad applicability.This method requires standard molecular biology equipment but does not rely on sequencing-based platforms.
Authors: Qile Gao, Ting Zhang, Yiqun Yuan, Gang Li, Bing Li, Chenglong Xiong
Institutions: Fudan University, Jinshan Hospital of Fudan University
Language: English
Subfield: Biomedical Engineering
Field: Engineering
Domain: Physical Sciences
Open Access status: bronze
OpenAlex ID: W4411502254