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Background  Recently, production of 16S rRNA methylases by Gram-negative bacilli has emerged as a novel mechanism for high-level resistance to aminoglycosides by these organisms in a variety of geographic locations. Therefore, the spread of high-level aminoglycoside resistance determinants has become a great concern.    Methods  Between January 2006 and July 2008, 680 distinct  Escherichia coli  clinical isolates were collected from a teaching hospital in Wenzhou, China. PCR and DNA sequencing were used to identify 16S rRNA methylase and extended-spectrum β-lactamase (ESBL) genes, including  armA  and  rmtB , and in situ hybridization was performed to determine the location of 16S rRNA methylase genes. Conjugation experiments were subsequently performed to determine whether aminoglycoside resistance was transferable from the  E. coli  isolates via 16S rRNA methylase-bearing plasmids. Homology of the isolates harboring 16S rRNA methylase genes was determined using pulse-field gel electrophoresis (PFGE).    Results  Among the 680  E. coli  isolates, 357 (52.5%), 346 (50.9%) and 44 (6.5%) isolates were resistant to gentamicin, tobramycin and amikacin, respectively. Thirty-seven of 44 amikacin-resistant isolates harbored 16S rRNA methylase genes, with 36 of 37 harboring the  rmtB  gene and only one harboring  armA . The positive rates of 16S rRNA methylase genes among all isolates and amikacin-resistant isolates were 5.4% (37/680) and 84.1% (37/44), respectively. Thirty-one isolates harboring 16S rRNA methylase genes also produced ESBLs. In addition, high-level aminoglycoside resistance could be transferred by conjugation from four  rmtB -positive donors. The plasmids of incompatibility groups IncF, IncK and IncN were detected in 34, 3 and 3 isolates, respectively. Upstream regions of the  armA  gene contained  IS CR1 and  tnpU , the latter a putative transposase gene,. Another putative transposase gene,  tnpD , was located within a region downstream of  armA . Moreover, a transposon, Tn 3 , was located upstream of the  rmtB . Nineteen clonal patterns were obtained by PFGE, with type H representing the prevailing pattern.    Conclusion  A high prevalence of plasmid-mediated  rmtB  gene was found among clinical  E. coli  isolates from a Chinese teaching hospital. Both horizontal gene transfer and clonal spread were responsible for the dissemination of the  rmtB  gene.

High prevalence of plasmid-mediated 16S rRNA methylase gene rmtB among Escherichia coliclinical isolates from a Chinese teaching hospital