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Molecular Diversity And Evolutionary Relationships Of Tn 1546 -Like Elements In Enterococci From Humans And Animals

Rob J. L. Willems, Janetta Top, Nicole van den Braak, Alex van Belkum, Dik J. Mevius, Giel Hendriks, Marga van Santen-Verheuvel, Jan D. A. van Embden

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ABSTRACT We report on a detailed study on the molecular diversity and evolutionary relationships of Tn 1546 -like elements in vancomycin-resistant enterococci (VRE) from humans and animals. Restriction fragment length polymorphism (RFLP) analysis of the VanA transposon of 97 VRE revealed seven different Tn 1546 types. Subsequent sequencing of the complete VanA transposons of 13 VRE isolates representing the seven RFLP types followed by sequencing of the identified polymorphic regions in 84 other VanA transposons resulted in the identification of 22 different Tn 1546 derivatives. Differences between the Tn 1546 types included point mutations in orf1 , vanS , vanA , vanX , and vanY . Moreover, insertions of an IS 1216V -IS 3 -like element in orf1 , of IS 1251 in the vanS-vanH intergenic region, and of IS 1216V in the vanX-vanY intergenic region were found. The presence of insertion sequence elements was often associated with deletions in Tn 1546 . Identical Tn 1546 types were found among isolates from humans and farm animals in The Netherlands, suggesting the sharing of a common vancomycin resistance gene pool. Application of the genetic analysis of Tn 1546 to VRE isolates causing infections in hospitals in Oxford, United Kingdom, and Chicago, Ill., suggested the possibility of the horizontal transmission of the vancomycin resistance transposon. The genetic diversity in Tn 1546 combined with epidemiological data suggest that the DNA polymorphism among Tn 1546 variants can successfully be exploited for the tracing of the routes of transmission of vancomycin resistance genes.