Genetic Rearrangements of Tn 1546 -Like Elements in Vancomycin-Resistant Enterococcus faecium Isolates Collected from Hospitalized Patients over a Seven-Year Period

Abstract

ABSTRACT The heterogeneity of Tn 1546 results from point mutations, deletions, and the integration of insertion sequence (IS) elements. Among these variations, the presence of IS elements accounts for much of the heterogeneity. Such a rearrangement could play a key role in the evolution of the vanA gene cluster, and hence, it may modify its transferability. In this study, we characterized the consequence of Tn 1546 in vanA -containing Enterococcus faecium isolates collected from patients over time. From 1998 to 2004, 57 vanA -containing E. faecium isolates were collected from hospitalized patients at Ajou University Hospital in Korea. PCR amplification of internal regions of Tn 1546 was performed, and both DNA strands were directly sequenced by the dideoxy termination method. All isolates were divided into three main types, including the prototype, according to the distribution of IS elements integrated into Tn 1546 elements. Type I was characterized by an IS 1542 insertion in the orf2-vanR intergenic region and an IS 1216V insertion in the vanX-vanY intergenic region. Type II was represented by the presence of two copies of IS 1216V at the 3′ end of IS 1542 and in the vanX-vanY intergenic region, as well as IS 1542 in the orf2-vanR intergenic region. Seventeen strains isolated from 1998 to 2000 represented type I, and 38 strains isolated from 2000 to 2004 represented type II. The remaining two isolates were the prototype. The tendency for the rearrangement of Tn 1546 was that the sequences were shortened as time passed, especially at the left or the right end, and hence, this could gradually modulate their transferability.