Identification and characterization of a novel incompatibility group X3 plasmid carrying bla_NDM-1 in Enterobacteriaceae isolates with epidemiological links to multiple geographical areas in China

Abstract

The New Delhi metallo-β-lactamase (NDM-1) is one of the most important resistance traits in Enterobacteriaceae. We characterized nine bla_NDM-1 producing Enterobacteriaceae recovered from seven patients who have recently travelled or been treated in India (n=1) or mainland China (n=6) during December 2010–May 2012. All the China-linked patients had no links to the Indian subcontinent. The bla_NDM-1 carrying plasmids belonged to the novel IncX3 (∼50 kb, in seven isolates including two Escherichia coli, two Klebsiella pneumoniae, one Citrobacter freundii, one Enterobacter aerogenes and one E. cloacae), IncA/C2 (∼140 kb, in one E. coli) or FII-F1B groups (∼110 kb, in one E. coli). Restriction fragment length polymorphism analysis of the seven IncX3 plasmids revealed identical pattern in six and two bands difference in the remaining one. The IncX3 plasmids carrying bla_NDM-1 were epidemiologically linked to Guangzhou (n=1), Hunan (n=4), Haifeng (n=1) and Dongguan (n=1) in mainland China. Complete sequencing of the IncX3 plasmid pNDM-HN380 revealed that it was 54 035 bp long and encoded 52 open reading frames. The bla_NDM-1 gene was found in a transposon-like structure flanked by ISAba125 and IS26, inserted into the plasmid genetic load region. The sequences of the bla_NDM-1 containing module within the two IS elements were identical to those previously described for bla_NDM-1-positive Tn125 in the plasmids or chromosome of Acinetobacter isolates. In summary, this is the first description of IncX3 plasmids carrying bla_NDM-1. The findings indicate the worrisome involvement of an epidemic plasmid in the dissemination of NDM-1 in China.

Keywords:

• antimicrobial resistance
• carbapenemases
• epidemiology
• incompatibility groups
• NDM-1
• plasmids

We thank Levina Lam, Ben Ho and Wilson Chan of the Centre for Genomic Sciences, University of Hong Kong for technical assistance. This work was supported by a commission grant (grant NO HK-09-01) from the Research Fund for the Control of Infectious Diseases (RFCID) of the Health and Food Bureau of the Hong Kong Special Administative Region Government. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.