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Preparation and application of microarrays for the detection of antibiotic resistance genes in samples isolated from Changchun, China

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Abstract

The emergence of antibiotic-resistant bacteria, especially tetracycline- and β-lactam-resistant bacteria, poses a great threat to human health. The purpose of this study was to develop and apply a suitable gene microarray for the detection of antibiotic resistance genes. We isolated 463 strains of bacteria from a hospital, a veterinary station, an animal nursery, and living environment of Changchun, China. After screening, it was found that 93.9% of these bacteria were resistant to tetracycline, 74.9% to ampicillin, 55.6% to deoxycycline, and 41.7% to ciprofloxacin. For amplification of antibiotic genes, we designed 28 pairs of primers. In addition, 28 hybridization probes for these genes were developed. The DNA microarray analysis was performed at 42°C for 5 h. We were successful in detecting 12 resistance genes by microarray analysis. After detection, we also evaluated the sensitivity of the microarray analysis. The LDL (Lowest Detection Level) of the microarray was 1 × 106 copies/ml of template DNA. It is believed that such microarray-based determination of tetracycline and β-lactam resistance genes can have a potential application in clinical studies in the future.

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Acknowledgments

This work was supported by a grant from the Development Program of the Science and Technology Department of Jilin Province. We would like to thank the Eleventh Institute of Academy of Military Medical Sciences of PLA for the support and providing instrumentation.

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Authors and Affiliations

  1. College of Animal Science and Veterinary Medicine, Jilin University, Changchun, Jilin, China

    Hao Lu, Yingchao Wang, Yuan Dang, Fuxian Zhang & Xin Feng

  2. The Eleventh Institute of Academy of Military Medical Sciences of PLA, Changchun, China

    Xinglong Wang, Xulong Lang, Jie Tang & Xiaoyan Li

Authors
  1. Hao Lu
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  2. Xinglong Wang
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  3. Xulong Lang
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  4. Yingchao Wang
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  5. Yuan Dang
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  6. Fuxian Zhang
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  7. Jie Tang
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  8. Xiaoyan Li
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  9. Xin Feng
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Correspondence to Xinglong Wang.

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Lu, H., Wang, X., Lang, X. et al. Preparation and application of microarrays for the detection of antibiotic resistance genes in samples isolated from Changchun, China. Mol Biol Rep 37, 1857–1865 (2010). https://doi.org/10.1007/s11033-009-9621-4

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  • DOI: https://doi.org/10.1007/s11033-009-9621-4

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