Rapid Detection of Antibiotic Resistance of Klebsiella pneumoniae by Denaturing High-performance Liquid Chromatography (DHPLc) / 中华医院感染学杂志

ABSTRACT

OBJECTIVE To type the genes of plasmid DNA in 54 clinical Klebsiella pneumoniae isolates producing extended -spectrum beta-lactamases (SHV) by denaturing high-performance liquid chromatography (DHPLc) and evaluate their sensitivity and specificity, and explore a rapid and convenient method for detecting the antibiotic resistance of K. pneumoniae. METHODS Plasmid DNA from each extended-spectrum beta-lactamase (SHV) producing strain was subjected to PCR amplification. After we performed DNA sequencing of these amplicons and identification of mutation and their genotype, DHPLc was undertaken to investigate whether its results correlate the distinctive chromatogram with each genotype. RESULTS All the strains were found abnormal elution peaks (two or three peaks) which were different from each other. The result of DNA sequencing demonstrated that all the strains had DNA mutation in comparison with SHV-1. Moreover, DHPLc could produce specific peak patterns that correlate with genotype. CONCLUSIONS The sensitivity of DHPLc is 100% in this study. And each genotype is corresponded to specific peak pattern. So we can use DHPLc technique to type the genes of plasmid DNA in K. pneumoniae and detect mutations rapidly. DHPLc not only has high accuracy , but also is a convenient and rapid technique for the detection of mutation in the bacterial genome. It has a great potential clinical value.