Oligonucleotide microarrays with horseradish peroxidase-based detection for the identification of extended-spectrum β-lactamases

Biosensors & Bioelectronics
Mayya Yu RubtsovaAlexey M Egorov

Abstract

Production of extended-spectrum β-lactamases (ESBLs) is the one of most widespread and clinically significant mechanism of Enterobacteriaceae resistance towards modern β-lactam antibiotics. There are known 400 ESBLs, with the majority represented by the enzymes of TEM, SHV and CTX-M families. Oligonucleotide microarrays with colorimetric detection have been developed for the purposes of determination of ESBLs and inhibitor-resistant β-lactamases using horseradish peroxidase (HRP). Specific oligonucleotide probes have been designed for the identification of β-lactamase family and important SNPs responsible for the broadening of substrate specificity and tolerance to inhibitors. Multiplex PCR has been developed for simultaneous amplification and labeling of full-size genes of TEM-, SHV- and CTX-M-type β-lactamases with biotin. The labeled target DNA is then hybridized with specific oligonucleotide probes immobilized on a porous membrane support. After hybridization, biotin-labeled DNA duplexes are stained with the streptavidin-HRP conjugate detected colorimetrically. Design of oligonucleotide probes and optimization of hybridization conditions ensure the specificity of all control ESBLs identification. The newly developed method ...Continue Reading

Citations

Jun 1, 2013·Biosensors & Bioelectronics·Shuang ZhouBin Xie
Feb 11, 2015·Journal of Dairy Science·Shuang ZhouYongning Wu
Apr 15, 2015·Biochemistry. Biokhimii︠a︡·V G GrigorenkoA M Egorov
Oct 5, 2013·Diagnostic Microbiology and Infectious Disease·Agnese LupoAndrea Endimiani
May 7, 2016·Analytical Chemistry·Yiping ChenXingyu Jiang

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