The purpose of this study was to describe the presence of the novel optrA gene among clinical isolates of enterococci in a Spanish teaching hospital (May 2016–April 2017). optrA and cfr genes were screened by PCR in all isolates showing linezolid minimal inhibitory concentration (MIC) ≥4 mg/L. The genetic relatedness of the isolates, the presence of resistance and virulence genes, and the genetic environment of optrA were assessed by whole-genome sequencing (WGS). Six of 1,640 enterococci had linezolid MIC ≥4 mg/L. Among them, the optrA gene was detected in five Enterococcus faecalis isolated from unrelated patients. Although none of them had received linezolid or chloramphenicol, all had antecedents of recent quinolone consumption. WGS analysis revealed the existence of two different genotypes: ST585 and ST474. cfr was not detected in any of the isolates. No mutations were detected among the 23S ribosomal RNA and the ribosomal proteins L3, L4, and L22. Both genotypes also carried genes related to aminoglycoside, lincosamide, macrolide, phenicol, and tetracycline resistance. Detection of optrA in a setting with low linezolid consumption and among patients without antecedents of oxazolidinone therapy is of concern.
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References
1.
TsiodrasS., GoldH.S., SakoulasG., EliopoulosG.M., WennerstenC., VenkataramanL., MoelleringR.C., and FerraroM.J.. 2001. Linezolid resistance in a clinical isolate of Staphylococcus aureus. Lancet., 358:207–208.
2.
GonzalesR.D., SchreckenbergerP.C., GrahamM.B., KelkarS., DenBestenK., and QuinnJ.P.. 2001. Infections due to vancomycin-resistant Enterococcus faecium resistant to linezolid. Lancet., 357:1179.
3.
MendesR.E., DeshpandeL.M., and JonesR.N.. 2014. Linezolid update: stable in vitro activity following more than a decade of clinical use and summary of associated resistance mechanisms. Drug Resist. Updat., 17:1–12.
4.
SchwarzS., WerckenthinC., and KehrenbergC.. 2000. Identification of a plasmid-borne chloramphenicol-florfenicol resistance gene in Staphylococcus sciuri. Antimicrob. Agents Chemother., 44:2530–2533.
5.
WangY., LvY., CaiJ., SchwarzS., CuiL., HuZ., ZhangR., LiJ., ZhaoQ., HeT., WangD., WangZ., ShenY., LiY., FesslerA.T., WuC., YuH., DengX., XiaX., and ShenJ.. 2015. A novel gene, optrA that confers transferable resistance to oxazolidinones and phenicols and its presence in Enterococcus faecalis and Enterococcus faecium of human and animal origin. J. Antimicrob. Chemother., 70:2182–2190.
6.
CaiJ., WangY., SchwarzS., LvH., LiY., LiaoK., YuS., ZhaoK., GuD., WangX., ZhangR., and ShenJ.. 2015. Enterococcal isolates carrying the novel oxazolidinone resistance gene optrA from hospitals in Zhejiang, Guangdong, and Henan, China, 2010–2014. Clin. Microbiol. Infect., 21:1095. e1091–e1094.
7.
BrencianiA., MorroniG., VincenziC., MansoE., MingoiaM., GiovanettiE., and VaraldoP.E.. 2016. Detection in Italy of two clinical Enterococcus faecium isolates carrying both the oxazolidinone and phenicol resistance gene optrA and a silent multiresistance gene cfr. J. Antimicrob. Chemother., 71:1118–1119.
8.
CavacoL.M., BernalJ.F., ZankariE., LeonM., HendriksenR.S., Perez-GutierrezE., AarestrupF.M., and Donado-GodoyP.. 2016. Detection of linezolid resistance due to the optrA gene in Enterococcus faecalis from poultry meat from the American continent (Colombia). J. Antimicrob. Chemother., 72:678–683.
9.
CuiL., WangY., LvY., WangS., SongY., LiY., LiuJ., XueF., YangW., and ZhangJ.. 2016. Nationwide surveillance of novel oxazolidinone resistance gene optrA in Enterococcus isolates in China from 2004 to 2014. Antimicrob. Agents Chemother., 60:7490–7493.
10.
HeT., ShenY., SchwarzS., CaiJ., LvY., LiJ., FesslerA.T., ZhangR., WuC., ShenJ., and WangY.. 2016. Genetic environment of the transferable oxazolidinone/phenicol resistance gene optrA in Enterococcus faecalis isolates of human and animal origin. J. Antimicrob. Chemother., 71:1466–1473.
11.
GawryszewskaI., ZabickaD., HryniewiczW., and SadowyE.. 2017. Linezolid-resistant enterococci in Polish hospitals: species, clonality and determinants of linezolid resistance. Eur. J. Clin. Microbiol. Infect. Dis., 36:1279–1286.
12.
MendesR.E., HoganP.A., JonesR.N., SaderH.S., and FlammR.K.. 2016. Surveillance for linezolid resistance via the Zyvox(R) Annual Appraisal of Potency and Spectrum (ZAAPS) programme (2014): evolving resistance mechanisms with stable susceptibility rates. J. Antimicrob. Chemother., 71:1860–1865.
13.
FanR., LiD., WangY., HeT., FesslerA.T., SchwarzS., and WuC.. 2016. Presence of the optrA gene in methicillin-resistant Staphylococcus sciuri of porcine origin. Antimicrob. Agents Chemother., 60:7200–7205.
14.
LiD., WangY., SchwarzS., CaiJ., FanR., LiJ., FesslerA.T., ZhangR., WuC., and ShenJ.. 2016. Co-location of the oxazolidinone resistance genes optrA and cfr on a multiresistance plasmid from Staphylococcus sciuri. J. Antimicrob. Chemother., 71:1474–1478.
15.
HuangJ., ChenL., WuZ., and WangL.. 2017. Retrospective analysis of genome sequences revealed the wide dissemination of optrA in Gram-positive bacteria. J. Antimicrob. Chemother., 72:614–616.
16.
SharkeyL.K., EdwardsT.A., and O'NeillA.J.. 2016. ABC-F proteins mediate antibiotic resistance through ribosomal protection. mBio., 7:e01975.
17.
SierraJ.M., CamoezM., TubauF., GaschO., PujolM., MartinR., and DomínguezM.A.. 2013. Low prevalence of Cfr-mediated linezolid resistance among methicillin-resistant Staphylococcus aureus. in a Spanish hospital: case report on linezolid resistance acquired during linezolid therapy. PLoS One. 8:e5, 9215.
18.
Clinical and Laboratory Standards Institute. 2017. Performance standards for antimicrobial susceptibility testing; 27th informational supplement. CLSI document M100-S27. CLSI, Wayne, PA.
19.
Ruiz-GarbajosaP., BontenM.J., RobinsonD.A., TopJ., NallapareddyS.R., TorresC., CoqueT.M., CantonR., BaqueroF., MurrayB.E., del CampoR., and WillemsR.J.. 2006. Multilocus sequence typing scheme for Enterococcus faecalis reveals hospital-adapted genetic complexes in a background of high rates of recombination. J. Clin. Microbiol., 44:2220–2228.
20.
YasufukuT., ShigemuraK., ShirakawaT., MatsumotoM., NakanoY., TanakaK., ArakawaS., KawabataM., and FujisawaM.. 2011. Mechanisms of and risk factors for fluoroquinolone resistance in clinical Enterococcus faecalis isolates from patients with urinary tract infections. J. Clin. Microbiol., 49:3912–3916.
21.
ZhaoQ., WangY., WangS., WangZ., DuX.D., JiangH., XiaX., ShenZ., DingS., WuC., ZhouB., WuY., and ShenJ.. 2016. Prevalence and abundance of florfenicol and linezolid resistance genes in soils adjacent to swine feedlots. Sci. Rep., 6:32192.
22.
FreitasA.R., ElghaiebH., Leon-SampedroR., AbbassiM.S., NovaisC., CoqueT.M., HassenA., and PeixeL.. 2017. Detection of optrA in the African continent (Tunisia) within a mosaic Enterococcus faecalis plasmid from urban wastewaters. J. Antimicrob. Chemother., 72:3245–3251.
23.
VorobievaV., RoerL., JustesenU.S., HansenF., Frimodt-MøllerN., HasmanH., and HammerumA.M.. 2017. Detection of the optrA gene in a clinical ST16 Enterococcus faecalis isolate in Denmark. J. Glob. Antimicrob. Resist., 10:12–13.
24.
MendesR.E., DeshpandeL., StreitJ.M., SaderH.S., CastanheiraM., HoganP.A., and FlammR.K.. 2018. ZAAPS programme results for 2016: an activity and spectrum analysis of linezolid using clinical isolates from medical centres in 42 countries. J. Antimicrob. Chemother., 73:1880–1887.
25.
FlammR.K., MendesR.E., HoganP.A., StreitJ.M., RossJ.E., and JonesR.N.. 2016. Linezolid surveillance results for the United States (LEADER surveillance program 2014). Antimicrob. Agents Chemother., 60:2273–2280.
26.
NseirS., Di PompeoC., SoubrierS., DelourP., LenciH., Roussel-DelvallezM., OnimusT., SaulnierF., MathieuD., and DurocherA.. 2005. First-generation fluoroquinolone use and subsequent emergence of multiple drug-resistant bacteria in the intensive care unit. Crit. Care Med., 33:283–289.
27.
MulanovichV.E., HubandM.D., MccurdyS.P., LemmonM.M., LescoeM., JiangY., RolstonK.V., and LaSalaP.R.. 2010. Emergence of linezolid-resistant coagulase-negative Staphylococcus in a cancer centre linked to increased linezolid utilization. 2001–2004. J. Antimicrob. Chemother., 65:2001–2004.
28.
FreitasA.R., NovaisC., Ruiz-GarbajosaP., CoqueT.M., and PeixeL.. 2009. Clonal expansion within clonal complex 2 and spread of vancomycin-resistant plasmids among different genetic lineages of Enterococcus faecalis from Portugal. J. Antimicrob. Chemother., 63:1104–1111.
29.
KawalecM., PietrasZ., DaniłowiczE., JakubczakA., GniadkowskiM., HryniewiczW., and WillemsR.J.. 2007. Clonal structure of Enterococcus faecalis isolated from Polish hospitals: characterization of epidemic clones. J. Clin. Microbiol., 45:147–153.
30.
Arredondo-AlonsoS., WillemsR.J., van SchaikW., and SchurchA.C.. 2017. On the (im)possibility of reconstructing plasmids from whole-genome short-read sequencing data. Microb. Genom., 3:e000128.
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