IS1542 is an insertion sequence-like element which was originally identified in the orf2-vanR intergenic region of VanA resistance elements of glycopeptide-resistant Enterococcus faecium from hospital patients and from non-human sources in the UK. The nucleotide sequence of IS1542 was determined. It showed 81% homology with IS256 and contained an open reading frame sufficient to encode a 390 amino acid peptide predicted to have 87.2% identity with the transposase of IS256. By PCR, IS1542 was detected in the genomes of 26 of 28 (93%) VanA enterococci isolated from poultry in the UK and Ireland, but in only two of 65 (3%) glycopeptide-sensitive or VanC enterococci from hospital patients in the UK and Brazil. IS1542 may be a useful marker for evolutionary and epidemiological studies of VanA glycopeptide resistance in enterococci.