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Journal of Clinical Microbiology, November 2007, p. 3514-3521, Vol. 45, No. 11
0095-1137/07/$08.00+0     doi:10.1128/JCM.02340-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Use of a DNA Microarray for Simultaneous Detection of Antibiotic Resistance Genes among Staphylococcal Clinical Isolates{triangledown}

Ling-Xiang Zhu,1,2,3 Zhi-Wei Zhang,3,4 Can Wang,3,4 Hua-Wei Yang,3,4 Di Jiang,3,4 Qiong Zhang,3,4 Keith Mitchelson,1,3,4 and Jing Cheng1,2,3,5*

Medical Systems Biology Research Center, Tsinghua University School of Medicine, Beijing 100084, China,1 Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China,2 National Engineering Research Center for Beijing Biochip Technology, 18 Life Science Parkway, Changping District, Beijing 102206, China,3 CapitalBio Corp., 18 Life Science Parkway, Changping District, Beijing 102206, China,4 State Key Laboratory of Biomembrane and Membrane Biotechnology, Tsinghua University, Beijing 100084, China5

Received 19 November 2006/ Returned for modification 22 January 2007/ Accepted 20 August 2007

We developed a multiplex asymmetric PCR (MAPCR)-based DNA microarray assay for characterization of the clinically relevant antibiotic resistance genes leading to penicillin, methicillin, aminoglycoside, macrolide, lincosamide, and streptogramin B (MLSB) resistance in staphylococci. The DNA-based assay involves detection of specific conserved regions of the mecA, blaZ (methicillin and penicillin resistance), aac(6')-Ie-aph(2''') (aminoglycoside resistance), ermA and ermC genes (MLSB resistance), and the msrA gene (macrolide and streptogramin B resistance). The microarray uses a variable sequence region of the 16S rRNA gene to broadly differentiate between Staphylococcus aureus and other coagulase-negative staphylococci (CoNS). The performance of the microarray was validated with a total of 178 clinically important S. aureus and 237 CoNS isolates, with correlations of 100% for S. aureus to CoNS discrimination and more than 90% for antibiotic resistance between the genotypic analysis determined by the microarray and the phenotype determined by standard methods of species identification and susceptibility testing. The major discrepant results were 17 mecA-positive CoNS and 60 aac(6')-Ie-aph(2''')-positive CoNS isolates measured by microarray that were susceptible to the corresponding antibiotics based on disk diffusion assay. Overall, this microarray-based assay offers a simultaneous, fast (≤5 h), and accurate identification of antibiotic resistance genes from a single colony, as well as species classification. Our extensive validation of the microarray suggests that it may be a useful tool to complement phenotypic susceptibility testing in clinical laboratories and to survey the spread of antibiotic resistance determinants in epidemiological studies.

* Corresponding author. Mailing address: Medical Systems Biology Research Center, Tsinghua University School of Medicine, Beijing 100084, China. Phone: (86)-10-62772239. Fax: (86)-10-80726898. E-mail: jcheng{at}tsinghua.edu.cn

{triangledown} Published ahead of print on 29 August 2007.

Journal of Clinical Microbiology, November 2007, p. 3514-3521, Vol. 45, No. 11
0095-1137/07/$08.00+0     doi:10.1128/JCM.02340-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Miller, M. B., Tang, Y.-W. (2009). Basic Concepts of Microarrays and Potential Applications in Clinical Microbiology. Clin. Microbiol. Rev. 22: 611-633 [Abstract] [Full Text]  


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