Affiliation:
1. Graduate School Hebei North University Zhangjiakou China
2. Department of Clinical Laboratory Hebei General Hospital Shijiazhuang China
3. National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention Beijing China
4. Graduate School Hebei Medical University Shijiazhuang China
5. Hebei Key Laboratory of Molecular Medicine Shijiazhuang China
6. Hebei Clinical Research Center for Laboratory Medicine Shijiazhuang China
Abstract
ABSTRACTObjectiveThis study aimed to establish a highly sensitive and rapid single‐tube, two‐stage, multiplex recombinase‐aided qPCR (mRAP) assay to specifically detect the khe, blaKPC‐2, and blaNDM‐1 genes in Klebsiella pneumoniae.MethodsmRAP was carried out in a qPCR instrument within 1 h. The analytical sensitivities of mRAP for khe, blaKPC‐2, and blaNDM‐1 genes were tested using recombinant plasmids and dilutions of reference strains. A total of 137 clinical isolates and 86 sputum samples were used to validate the clinical performance of mRAP.ResultsmRAP achieved the sensitivities of 10, 8, and 14 copies/reaction for khe, blaKPC‐2, and blaNDM‐1 genes, respectively, superior to qPCR. The Kappa value of qPCR and mRAP for detecting khe, blaKPC‐2, and blaNDM‐1 genes was 1, 0.855, and 1, respectively (p < 0.05).ConclusionmRAP is a rapid and highly sensitive assay for potential clinical identification of khe, blaKPC‐2, and blaNDM‐1 genes in K. pneumoniae.
Funder
National Key Research and Development Program of China
National Natural Science Foundation of China
Cited by
1 articles.
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