AI Article Synopsis
- - Increased use of colistin in medicine has led to the emergence of resistance genes, which this study aims to detect using a real-time PCR assay with TaqMan probe-based chemistry.
- - The assay demonstrated high amplification efficiency (98% and 96%) for specific resistance genes, and was able to detect as few as 10 cfu/mL of bacteria without cross-reacting with other resistance genes.
- - With a 100% sensitivity and specificity confirmed through validation with characterized bacterial isolates, this assay serves as a rapid and precise tool for identifying colistin resistance in laboratory settings, meeting CLIA quality standards.
Article Abstract
Increased use of colistin in both human and veterinary medicine has led to the emergence of plasmid-mediated colistin resistance ( genes). In this study, we report the development of a real-time PCR assay using TaqMan probe-based chemistry for detection of genes from bacterial isolates. Positive control isolates harboring and yielded exponential amplification curves with the assay, and the amplification efficiency was 98% and 96% for and , respectively. Each target gene could be reproducibly detected from a sample containing 10 cfu/mL of -harboring bacteria, and there was no cross-reactivity with DNA extracted from several multidrug-resistant bacteria harboring other resistance genes, but lacking genes. Both sensitivity and specificity of the real-time PCR assay were 100% in a method validation performed with a set of 25 previously well-characterized bacterial isolates containing -positive and -negative bacteria. This newly developed assay is a rapid and sensitive tool for detecting emerging genes in cultured bacterial isolates. The assay was successfully validated according to quality standards of the Clinical Laboratory Improvement Amendments (CLIA).
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| http://dx.doi.org/10.1089/mdr.2018.0417 | DOI Listing |
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