ABSTRACT
OBJECTIVE: This 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. METHODS: mRAP 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. RESULTS: mRAP 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). CONCLUSION: mRAP is a rapid and highly sensitive assay for potential clinical identification of khe, blaKPC-2, and blaNDM-1 genes in K. pneumoniae.
Subject(s)
Klebsiella pneumoniae , Multiplex Polymerase Chain Reaction , beta-Lactamases , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/isolation & purification , beta-Lactamases/genetics , Humans , Multiplex Polymerase Chain Reaction/methods , Klebsiella Infections/microbiology , Klebsiella Infections/diagnosis , Sensitivity and Specificity , Real-Time Polymerase Chain Reaction/methods , Bacterial Proteins/genetics , Recombinases/genetics , Recombinases/metabolismABSTRACT
Conversion of cellulose to 1-(furan-2-yl)-2-hydroxyethanone has been demonstrated in concentrated zinc chloride solution under microwave irradiation. Compared with the conventional oil-bath heating mode, microwave irradiation significantly reduced the reaction time and increased the yield of 1-(furan-2-yl)-2-hydroxyethanone. A typical degradation reaction with cellulose produced 1-(furan-2-yl)-2-hydroxyethanone in 12.0% molar yield in ZnCl(2) solution (ZnCl(2)-H(2)O ratio=2.25:1, w/w) with microwave irradiation at 600 W for 5 minutes at 135°C.
