TY - JOUR
T1 - A classification system for plasmids from Enterococci and other Gram-positive bacteria
AU - Jensen, Lars Bogø
AU - Garcia-Migura, Lourdes
AU - Valenzuela, Antonio Jesus Sanchez
AU - Løhr, Mille
AU - Hasman, Henrik
AU - Aarestrup, Frank Møller
PY - 2010
Y1 - 2010
N2 - A classification system for plasmids isolated from enterococci and other Gram-positive bacteria was developed based on 111 published plasmid sequences from enterococci and other Gram-positive bacteria; mostly staphylococci. Based on PCR amplification of conserved areas of the replication initiating genes (rep), alignment of these sequences and using a cut off value of 80% identity on both protein and DNA level, 19 replicon families (rep-families) were defined together with several unique sequences. The prevalence of these rep-families was tested on 79 enterococcal isolates from a collection of isolates of animal and human origin. Difference in prevalence of the designed rep-families were detected with rep9 being most prevalent in Enterococcus faecalis and rep2 in Enterococcus faecium. In 33 % of the tested E. faecium and 32% of the tested E. faecalis no positive amplicons were detected. Furthermore, conjugation experiments were performed obtaining 30 transconjugants when selecting for antimicrobial resistance. Among them 19 gave no positive amplicons indicating presence of rep-families not tested for in this experimental setup.
AB - A classification system for plasmids isolated from enterococci and other Gram-positive bacteria was developed based on 111 published plasmid sequences from enterococci and other Gram-positive bacteria; mostly staphylococci. Based on PCR amplification of conserved areas of the replication initiating genes (rep), alignment of these sequences and using a cut off value of 80% identity on both protein and DNA level, 19 replicon families (rep-families) were defined together with several unique sequences. The prevalence of these rep-families was tested on 79 enterococcal isolates from a collection of isolates of animal and human origin. Difference in prevalence of the designed rep-families were detected with rep9 being most prevalent in Enterococcus faecalis and rep2 in Enterococcus faecium. In 33 % of the tested E. faecium and 32% of the tested E. faecalis no positive amplicons were detected. Furthermore, conjugation experiments were performed obtaining 30 transconjugants when selecting for antimicrobial resistance. Among them 19 gave no positive amplicons indicating presence of rep-families not tested for in this experimental setup.
KW - Plasmid typing
KW - PCR-based
KW - Gram-positive bacteria
KW - Enterococci
KW - Multiplex PCR
U2 - 10.1016/j.mimet.2009.10.012
DO - 10.1016/j.mimet.2009.10.012
M3 - Journal article
SN - 0167-7012
VL - 80
SP - 25
EP - 43
JO - Journal of Microbiological Methods
JF - Journal of Microbiological Methods
IS - 1
ER -