Microarray-based detection of 90 antibiotic resistance genes of gram-positive bacteria

Perreten, Vincent; Vorlet-Fawer, Lorianne; Slickers, Peter; Ehricht, Ralf; Kuhnert, Peter; Frey, Joachim (2005). Microarray-based detection of 90 antibiotic resistance genes of gram-positive bacteria. Journal of clinical microbiology, 43(5), pp. 2291-2302. Washington, D.C.: American Society for Microbiology 10.1128/JCM.43.5.2291-2302.2005

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A disposable microarray was developed for detection of up to 90 antibiotic resistance genes in gram-positive bacteria by hybridization. Each antibiotic resistance gene is represented by two specific oligonucleotides chosen from consensus sequences of gene families, except for nine genes for which only one specific oligonucleotide could be developed. A total of 137 oligonucleotides (26 to 33 nucleotides in length with similar physicochemical parameters) were spotted onto the microarray. The microarrays (ArrayTubes) were hybridized with 36 strains carrying specific antibiotic resistance genes that allowed testing of the sensitivity and specificity of 125 oligonucleotides. Among these were well-characterized multidrug-resistant strains of Enterococcus faecalis, Enterococcus faecium, and Lactococcus lactis and an avirulent strain of Bacillus anthracis harboring the broad-host-range resistance plasmid pRE25. Analysis of two multidrug-resistant field strains allowed the detection of 12 different antibiotic resistance genes in a Staphylococcus haemolyticus strain isolated from mastitis milk and 6 resistance genes in a Clostridium perfringens strain isolated from a calf. In both cases, the microarray genotyping corresponded to the phenotype of the strains. The ArrayTube platform presents the advantage of rapidly screening bacteria for the presence of antibiotic resistance genes known in gram-positive bacteria. This technology has a large potential for applications in basic research, food safety, and surveillance programs for antimicrobial resistance.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology

UniBE Contributor:

Perreten, Vincent; Kuhnert, Peter and Frey, Joachim

Subjects:

500 Science
500 Science > 570 Life sciences; biology
600 Technology > 630 Agriculture

ISSN:

0095-1137

Publisher:

American Society for Microbiology

Language:

English

Submitter:

Peter Kuhnert-Ryser

Date Deposited:

26 Jan 2014 18:05

Last Modified:

06 Dec 2014 02:08

Publisher DOI:

10.1128/JCM.43.5.2291-2302.2005

PubMed ID:

15872258

BORIS DOI:

10.7892/boris.39089

URI:

https://boris.unibe.ch/id/eprint/39089

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