Rapid Electrochemical Detection of New Delhi Metallo-beta-lactamase Genes To Enable Point-of-Care Testing of Carbapenem-Resistant Enterobacteriaceae

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• 作者：Jimmy Ming-Yuan Huang ; Grace Henihan ; Daniel Macdonald ; Annette Michalowski ; Kate Templeton ; Alan P. Gibb ; Holger Schulze ; Till T. Bachmann
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：August 4, 2015
• 年：2015
• 卷：87
• 期：15
• 页码：7738-7745
• 全文大小：342K
• ISSN：1520-6882

文摘

The alarming rate at which antibiotic resistance is occurring in human pathogens causes a pressing need for improved diagnostic technologies aimed at rapid detection and point-of-care testing to support quick decision making regarding antibiotic therapy and patient management. Here, we report the successful development of an electrochemical biosensor to detect bla_NDM, the gene encoding the emerging New Delhi metallo-beta-lactamase, using label-free electrochemical impedance spectroscopy (EIS). The presence of this gene is of critical concern because organisms harboring bla_NDM tend to be multiresistant, leaving very few treatment options. For the EIS assay, we used a bla_NDM-specific PNA probe that was designed by applying a new approach that combines in silico probe design and fluorescence-based DNA microarray validation with electrochemical testing on gold screen-printed electrodes. The assay was successfully demonstrated for synthetic targets (LOD = 10 nM), PCR products (LOD = 100 pM), and direct, amplification-free detection from a bla_NDM-harboring plasmid. The biosensor鈥檚 specificity, preanalytical requirements, and performance under ambient conditions were demonstrated and successfully proved its suitability for further point-of-care test development.