African Journal of Bacteriology Research
Subscribe to JBR
Full Name*
Email Address*

Article Number - E6F5F6C10030


Vol.5(6), pp. 68-75 , September 2013
DOI: 10.5897/JBR2013.0120
ISSN: 2006-9871



Full Length Research Paper

DNA microarray-based detection of antibiotic resistance genes of human isolates of Escherichia coli in Nigeria


Chijioke A. Nsofor1,2*, Christian U. Iroegbu1, Douglas R. Call2 and Margaret A. Davies2




 

1Department of Microbiology, University of Nigeria Nsukka, Enugu State, Nigeria.

2Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, USA.


Email: nsoforac@gmail.com.






 Accepted: 07 September 2013  Published: 30 September 2013

Copyright © 2013 Author(s) retain the copyright of this article.
This article is published under the terms of the Creative Commons Attribution License 4.0


 

Molecular epidemiology of human ecovariants of Escherichia coli from different regions of Nigeria were studied using their antibiotic susceptibility patterns, plasmid profile and DNA microarray techniques. E. coli was isolated using Eosin Methylene Blue agar (EMB) and identified by conventional microbiological technique. The isolates were tested against 14 antibiotics using the disc diffusion method. Genotyping was done using DNA microarray. Overall, 42 different antibiotics resistance clusters were observed, with each isolate showing resistance to at least four or more drugs tested. Of the 60 isolates genotyped with DNA microarray, 57 were identified as having, at least, one antimicrobial resistance gene. Among the 90 antibiotic resistance genes detected, bla-CMY-2 was the most prevalent occurring in 38 (63.3%) of the isolates. Other highly prevalent genes occurring in the human isolates, include strA 28(70%) aadE 28(70%); TEM1 11(27.5%); Sul2 14(35%); andTetA 21(52.5%). The microarray genotyping corresponded with the phenotype of the strains. Presence of drug-resistance genes/plasmids in commensal strains isolated from apparently healthy individuals is of great public health importance.

 

Key words: DNA microarray, E. coli, Nigeria.

Barlow M, Reik RA, Jacobs SD, Medina M, and Meyer MP. (2008). High rate of mobilization for blaCTX-Ms. Emerg. Infect. Dis. 14:423-428.
http://dx.doi.org/10.3201/eid1403.070405
PMid:18325257 PMCid:PMC2570810
 
Call DR, Chandler DP, Brockman F (2001). Fabrication of DNA microarrays using unmodified oligonucleotide probes. BioTech. 30:368-379.
 
Call DR, Bakko MK, Krug MJ, Roberts MC (2003a). Identifying antimicrobial resistance genes with DNA microarrays. Antimicrob. Agents Chemother. 47:3290-3295.
http://dx.doi.org/10.1128/AAC.47.10.3290-3295.2003
PMid:14506043 PMCid:PMC201141
 
Call DR, Borucki MK, Loge FJ (2003b). Detection of bacterial pathogens in environmental samples using DNA microarrays. J. Microbiol. Methods 53:235–243.
http://dx.doi.org/10.1016/S0167-7012(03)00027-7
 
Cheesbrough M (2000). District Laboratory Practice in Tropical Countries, Part 2. Cambridge University Press, Cambridge, UK; pp.434.
 
Cassone M, Marco MD, Francesco I, Marco RO, Gian MR, Gianni P (2006). DNA Microarray for Detection of Macrolides Resistance Genes. Antimicrob. Agents Chemother. pp. 2038-2041.
http://dx.doi.org/10.1128/AAC.01574-05
PMid:16723563 PMCid:PMC1479117
 
Clinical Laboratory Standards Institute (2006). Performance standards for Antimicrobial susceptibility testing. National committee for clinical laboratory standards, Wayne pa.
 
Martineau F, Picard FJ, Grenier L, Roy PH, Ouellette M, Bergeron MG (2000a). Multiplex PCR assays for the detection of clinically relevant antibiotic resistance genes in Staphylococci isolated from patients infected after cardiac surgery. J. Antimicrob. Chemother. 46:527-533.
http://dx.doi.org/10.1093/jac/46.4.527
PMid:11020248
 
Martineau F, Picard FJ, Grenier L, Roy PH, Ouellette M, Bergeron MG (2000b). Correlation between the resistance genotype determined by multiplex PCR assays and the antibiotic susceptibility patterns of Staphylococcus aureus and Staphylococcus epidermidis. Antimicrob. Agents Chemother. 44:231-238.
http://dx.doi.org/10.1128/AAC.44.2.231-238.2000
PMid:10639342 PMCid:PMC89663
 
Murakami S, Nakashima R, Yamashita E, Yamaguchi A (2002).Crystal structure of bacterial multidrug efflux transporter AcrB. Nature 419:587-593.
http://dx.doi.org/10.1038/nature01050
PMid:12374972
 
Nsofor CA, Iroegbu CU (2013) Antibiotic Resistance Profile of Escherichia coli Isolated from Five Major Geopolitical Zones of Nigeria. J. Bacteriol. Res. 5(3):29-34.
http://dx.doi.org/10.5897/JBR2012.035
 
Panicker G, Call DR, Krug MJ, Bej AK (2004). Detection of Pathogenic Vibrio spp. in Shellfish by Using Multiplex PCR and DNA Microarrays. Appl. Environ. Microbiol. pp.7436-7444.
http://dx.doi.org/10.1128/AEM.70.12.7436-7444.2004
PMid:15574946 PMCid:PMC535186
 
Perreten VL, Vorlet-Fawer P, Slickers R, Ehricht PK, Frey J (2005). Microarray-based detection of 90 antibiotic resistance genes of gram-positive bacteria. J. Clin. Microbiol. 43:2291-2302.
http://dx.doi.org/10.1128/JCM.43.5.2291-2302.2005
PMid:15872258 PMCid:PMC1153730
 
Peterson G, Jianfa B, Nagaraja TG, Sanjeev N (2010). Diagnostic microarray for human and animal bacterial diseases and their virulence and antimicrobial resistance genes. J. Microbiol. Methods 80:223-230.
http://dx.doi.org/10.1016/j.mimet.2009.12.010
PMid:20035807
 
Ye Y, He X, Szewczyk P, Nguyen T, Chang G (2006). Structure of the multidrug transporter EmrD from Escherichia coli. Sci. 312:741–744.
http://dx.doi.org/10.1126/science.1125629
PMid:16675700 PMCid:PMC3152482

 


APA (2013). DNA microarray-based detection of antibiotic resistance genes of human isolates of Escherichia coli in Nigeria. African Journal of Bacteriology Research, 5(6), 68-75.
Chicago Chijioke A. Nsofor, Christian U. Iroegbu, Douglas R. Call and Margaret A. Davies. "DNA microarray-based detection of antibiotic resistance genes of human isolates of Escherichia coli in Nigeria." African Journal of Bacteriology Research 5, no. 6 (2013): 68-75.
MLA Chijioke A. Nsofor, et al. "DNA microarray-based detection of antibiotic resistance genes of human isolates of Escherichia coli in Nigeria." African Journal of Bacteriology Research 5.6 (2013): 68-75.
   
DOI 10.5897/JBR2013.0120
URL http://academicjournals.org/journal/JBR/article-abstract/E6F5F6C10030

Subscription Form