Comprehensive Antibiotic Resistance Profiling

The bla_TEM, bla_SHV, bla_OXA-1-like and bla_CTX-M (group 1, 2 and 9) genes were detected via M-PCRs, and the bla_CTX-M group 8/25 was detected using a single PCR [50 (link),51 (link)]. The presence of plasmid-mediated AmpC β-lactamase genes, including ACC, FOX, MOX, DHA, CIT and EBC groups, was examined via M-PCR [50 (link)]. The carbapenem-resistant isolates were included for M-PCRs for the detection of carbapenemase-encoding genes [52 (link)]. PMQR genes, including aac(6′)-Ib-cr, qnrA, qnrB, qnrC, qnrD, qnrS and qepA, and a plasmid-mediated azithromycin resistance gene mph(A) were detected using PCRs [53 (link),54 (link),55 (link),56 (link),57 (link),58 (link),59 (link)]. The mcr-1 to -9 genes were screened in the colistin-resistant isolates using M-PCRs [60 (link),61 (link)]. The bla_CTX-M, bla_NDM and mcr variants were identified using nucleotide sequencing. All PCR reactions were carried out in 25 μL of reaction mixture including 5X Firepol^® Master Mix Ready-to-Load (Solis Biodyne, Tartu, Estonia), 0.2 µM of each primer and 1 μL of DNA template. The primers used for the detection of antimicrobial resistance genes are listed in Supplementary Table S2.

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Nittayasut N., Yindee J., Boonkham P., Yata T., Suanpairintr N, & Chanchaithong P. (2021). Multiple and High-Risk Clones of Extended-Spectrum Cephalosporin-Resistant and blaNDM-5-Harbouring Uropathogenic Escherichia coli from Cats and Dogs in Thailand. Antibiotics, 10(11), 1374.

Publication 2021

5X Firepol® Master Mix Ready-to-Load

Ampc β lactamase Antimicrobial Azithromycin Carbapenem Carbapenemase Colistin Genes Plasmid Primers

Corresponding Organization : Chulalongkorn University

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Variable analysis

independent variables

Detection of bla genes using M-PCR
Detection of plasmid-mediated AmpC β-lactamase genes using M-PCR
Detection of carbapenemase-encoding genes using M-PCR
Detection of PMQR genes and mph(A) gene using PCR
Screening of mcr-1 to -9 genes using M-PCR

dependent variables

Presence/absence of bla genes (bla_TEM, bla_SHV, bla_OXA-1-like, bla_CTX-M groups 1, 2, 9, and 8/25)
Presence/absence of plasmid-mediated AmpC β-lactamase genes (ACC, FOX, MOX, DHA, CIT, EBC groups)
Presence/absence of carbapenemase-encoding genes
Presence/absence of PMQR genes (aac(6')-Ib-cr, qnrA, qnrB, qnrC, qnrD, qnrS, qepA) and mph(A) gene
Presence/absence of mcr-1 to -9 genes

control variables

Volume of reaction mixture (25 μL)
Concentration of primers (0.2 μM)
Volume of DNA template (1 μL)
Positive and negative controls not explicitly mentioned

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