Molecular analysis of linezolid resistance in clinical Enterococcus faecium isolates by polymerase chain reaction and pyrosequencing.
Resistance to linezolid has been associated with a G2576T mutation in the 23 S rRNA gene. Clinical isolates of linezolid-sensitive and linezolid-resistant vancomycin-resistant Enterococcus faecium of a liver transplant patient have been analysed for the G2576T mutation by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP), conventional sequencing and pyrosequencing. A clear association between the number of mutated 23 S rRNA genes and the level of linezolid resistance has been demonstrated. Linezolid susceptibility re-emerged after cessation of linezolid therapy; however, the re-initiation of linezolid therapy resulted in the re-emergence of linezolid-resistant strains. Pyrosequencing rapidly detected the number of mutated alleles and is superior to conventional PCR-RFLP for the detection of heterozygous mutations.
Schnitzler
P
P
Department of Infectious Diseases, Virology, University of Heidelberg, Heidelberg, Germany.
Schulz
K
K
Lampson
C
C
Geiss
M
M
Geiss
H K
HK
D000081:Acetamides / Q000494:pharmacology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D016984:Enterococcus faecium / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D000069349:Linezolid; D023303:Oxazolidinones / Q000494:pharmacology*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D017422:Sequence Analysis, DNA
D000081:Acetamides; D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D023303:Oxazolidinones; D012338:RNA, Ribosomal, 23S; D000069349:Linezolid
Antimicrob Agents Chemother. 2003 Nov;47(11):3620-2
14576130
Antimicrob Agents Chemother. 2001 Jul;45(7):2154-6
11408243
Nucleic Acids Res. 2002 Apr 1;30(7):e31
11917037
J Clin Microbiol. 2002 Nov;40(11):4298-300
12409414
Antimicrob Agents Chemother. 2003 Oct;47(10):3318-20
14506047
Antimicrob Agents Chemother. 2002 Apr;46(4):1080-5
11897593
J Antimicrob Chemother. 2005 Oct;56(4):787-9
16143710
J Antimicrob Chemother. 2007 Sep;60(3):649-51
17623697
Antimicrob Agents Chemother. 2002 Oct;46(10):3334-6
12234875
J Antimicrob Chemother. 2002 Nov;50(5):743-6
12407134
Clin Infect Dis. 2002 Nov 15;35(10):1269-72
12410489
J Med Microbiol. 2010 Jun;59(Pt 6):672-678
20185550
Lancet. 2001 Apr 14;357(9263):1179
11323048
Eur J Clin Microbiol Infect Dis. 2006 Mar;25(3):197-8
16498524
Eur J Clin Microbiol Infect Dis. 2008 Sep;27(9):873-8
18421487
Antimicrob Agents Chemother. 1997 Oct;41(10):2132-6
9333037
J Antimicrob Chemother. 2004 Oct;54(4):818-20
15347637
Antimicrob Agents Chemother. 2008 Jun;52(6):2256-9
18391028