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Antimicrobial Agents and Chemotherapy, June 2006, p. 2038-2041, Vol. 50, No. 6
0066-4804/06/$08.00+0     doi:10.1128/AAC.01574-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

DNA Microarray for Detection of Macrolide Resistance Genes

Marco Cassone,1 Marco M. D'Andrea,2 Francesco Iannelli,1 Marco R. Oggioni,1 Gian Maria Rossolini,2 and Gianni Pozzi1*

LAMMB,1 FIBIM, Sezione di Microbiologia, Dipartimento di Biologia Molecolare, Università di Siena, Siena, Italy2

Received 10 December 2005/ Returned for modification 26 January 2006/ Accepted 31 March 2006

A DNA microarray was developed to detect bacterial genes conferring resistance to macrolides and related antibiotics. A database containing 65 nonredundant genes selected from publicly available DNA sequences was constructed and used to design 100 oligonucleotide probes that could specifically detect and discriminate all 65 genes. Probes were spotted on a glass slide, and the array was reacted with DNA templates extracted from 20 reference strains of eight different bacterial species (Streptococcus pneumoniae, Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Staphylococcus haemolyticus, Escherichia coli, and Bacteroides fragilis) known to harbor 29 different macrolide resistance genes. Hybridization results showed that probes reacted with, and only with, the expected DNA templates and allowed discovery of three unexpected genes, including msr(SA) in B. fragilis, an efflux gene that has not yet been described for gram-negative bacteria.

* Corresponding author. Mailing address: LAMMB, Università di Siena, Policlinico Le Scotte/V Lotto, Viale Bracci, 53100 Siena, Italy. Phone: 39 0577 233299. Fax: 39 0577 233334. E-mail: pozzi{at}unisi.it.

Antimicrobial Agents and Chemotherapy, June 2006, p. 2038-2041, Vol. 50, No. 6
0066-4804/06/$08.00+0     doi:10.1128/AAC.01574-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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