Aims: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important pathogens in the hospital environment. Monitoring of this pathogen by molecular characterization and phenotypic methods is important for the development of suitable infection control measures and proper therapy design. In this study, our aim was to investigate the molecular epidemiological characteristics of MRSA bloodstream isolates obtained from patients hospitalized at Ankara University Ibn-i Sina Hospital in a 10-year period (2002–2012) and monitor the possible changes. A total of 134 isolates were characterized according to their antimicrobial susceptibility profiles, biofilm formation capabilities, accessory gene regulator (agr) locus types, presence of genes encoding Panton-Valentine leukocidin (PVL), staphylococcal enterotoxins A–J (SEs A–J), toxic shock syndrome toxin, sasX, and genes associated with biofilm formation (icaD, icaA, IS256) by polymerase chain reaction. The staphylococcal cassette chromosome mec (SCCmec) types of isolates were also defined and their clonal relationships were investigated by pulsed-field gel electrophoresis (PFGE) analysis and multilocus sequence typing was performed for representative isolates obtained by PFGE. Results: The majority of the isolates were resistant to rifampin (100%), ciprofloxacin (97%), tetracycline (97.7%), and gentamicin (94.7%); 100% carried type-III SCCmec and 89.5% were agr type-1. All the isolates were negative for PVL, and sasX genes while all of them carried the icaD, icaA, and IS256 genes. The most common SE was enterotoxin A (97%). Four major PFGE patterns with the dominance of one pattern and seven unique patterns were obtained. All the representative PFGE isolates (n = 11) belonged to sequence type 239. Conclusion: We have documented the characteristics of the dominant MRSA clone in our hospital, which was a PVL (−), sasX (−) ST239 clone carrying sea (+) with type-III SCCmec, and type-1 agr locus.
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References
1.
EnrightM.C., DayN.P.J., DaviesC.E., PeacockS.J., and SprattB.G.2000. Multilocus sequence typing for characterization of methicillin-resistant and methicillin susceptible clones of Staphylococcus aureus. J. Clin. Microbiol., 38:1008–1015.
2.
OliveiraD.C., TomaszA., and de LencastreH.2002. Secrets of success of a human pathogen; molecular evolution of pandemic clones of methicillin resistant Staphylococcus aureus. Lancet Infect. Dis., 2:180–189.
3.
Echanis-AvilesG., Velazquez-MezaM.E., Aires-de-SousaM., Morfin-OteroR., Rodriguez-NoriegaE., Carnala-BarajasN., Esparza-AhumadaS., and de-LancastreH.2005. Molecular characterization of a dominant methicillin-resistant Staphylococcus aureus (MRSA) clone in a Mexican hospital (1999–2003). Clin. Microbiol. Infect., 12:22–28.
4.
EnrightM.C.D., RobinsonD.A., RandleG., FeilE.J., GrundmannH., and SprattB.G.. 2002. The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA) Proc. Natl. Acad. Sci. U.S.A., 99:7687–7692.
5.
GilletY., IssartelB., VanhemsP., FournetJ.C., LinaG., BesM., VandeneschF., YvesP., BrousseN, FloretD., and JeromeE.2002. Association between Staphylococcus aureus strains carrying the gene for the Panton-Valentine leukocidin and highly lethal necrotizing pneumonia in young immunocompetent patients. Lancet, 359:753–759.
6.
ItoT., KatayamaY., AsadaK., MoriN., TstutsumimotoN., and TiensasitornC.2001. Structural comparison of three types of staphylococcal cassette chromosome mec integrated in the chromosome in methicillin resistant Staphylococcus aureus. Antimicrob. Agents Chemother., 45:1323–1336.
7.
JarraudS., MougelC., ThioulouseJ., LinaG., MeugnierH., ForeyF., NesmeX., EtienneJ., and VandeneschF.2002. Relationships between Staphylococcus aureus genetic background, virulence factors, agr groups (alleles), and human disease. Infect. Immun., 70:631–641.
8.
LinaG., PiemontY., Godail-GamotF., BesM., PeterM.O., GauduchonV., VandeneschF., and EtienneJ.1999. Involvement of Panton-Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clin. Infect. Dis., 29:1128–1132.
9.
LiM., DuX., VillaruzA.E., DiepB.A., WangD., and SongY.2012. MRSA epidemic linked to a quickly spreading colonization and virulence determinant. Nat. Med., 18:816–819.
10.
AygenB., YörükA., YıldızO., AlpE., KocagözS., SümerkanB., and DoğanayM.2004. Bloodstream infections caused by Staphylococcus aureus in a university hospital in Turkey: clinical and molecular epidemiology of methicillin-resistant Staphylococcus aureus. Clin. Microbiol. Infect., 10:309–314.
11.
BorgM.A., de KrakerM., SciclunaE., van de Sande-BruinsmaN., TiemersmaE., MonenJ., GrundmannH., and ARMed Project Members and Collaborators. 2007. Prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in invasive isolates from southern and eastern Mediterranean countries. J. Antimicrob. Chemother., 60:1310–1315.
12.
AlpE., KlaassenC.H.W., DoganayM., AltoparlakU., AydinK., EnginA., KuzucuC., OzakinC., OzinelM.A., TurhanO., and VossA.2009. MRSA genotypes in Turkey: persistence over 10 years of a single clone of ST239. J. Infect. Dis., 58:433–438.
National Committee for Clinical and Laboratory Standards. 2013. Performance standards for antimicrobial susceptibility testing. Twenty-third Informational Supplement M100-S23. CLSI, Wayne, PA.
15.
ArajG.F., TalhoukR.S., SimanC.J., and MaasadM.J.1999. Discrepancies between mecA PCR and conventional tests used for detection of methicillin resistant Staphylococcus aureus. Int. J. Antimicrob. Agents, 11:47–52.
16.
KondoY., ItoT., MaX.X., WatanabeS., KreiswirthB.N., EtienneJ., and HiramatsuK.2007. Combination of multiplex PCRs for staphylococcal cassette chromosome mec type assignment: rapid identification system for mec, ccr, and major differences in junkyard regions. Antimicrob. Agents Chem., 51:264–274.
17.
GilotP., LinaG., CochardT., and PoutrelB.2002. Analysis of the genetic variability of genes encoding the RNA III-activating components agr and trap in a population of Staphylococcus aureus strains isolated from cows and mastitis. J. Clin. Microbiol., 40:4060–4067.
18.
MondayR.S., and BonachG.A.1999. Use of Multiplex PCR to detect classical and newly described pyrogenic toxin genes in Staphylococcal isolates. J. Clin. Microbiol., 37:3411–3414.
19.
MontanaroL., CampocciaD., PiriniV., RavaioliS., OttoM., and ArciolaC.R.2007. Antibiotic multiresistance strictly associated with IS256 and ica genes in Staphylococcus epidermidis strains from implant orthopedic infections. J. Biomed. Mater. Res. A, 83:813–818.
20.
VasudevanP., NairM.K., AnnamalaiT., and VenkitanaravananK.S.2003. Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation. Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation. Vet. Microbiol., 92:179–185.
21.
MulveyM.R., ChuiL., IsmailJ., LouieL., MurphyC., ChangN., AlfaM., and the Canadian Committee for the Standardization of Molecular Methods. 2001. Development of a Canadian standardized protocol for subtyping methicillin-resistant Staphylococcus aureus using pulsed-field gel electrophoresis. J. Clin. Microbiol., 39:3481–3485.
22.
CoombsG.W., NimmoG.R., BellJ.M., HuygensF., O'BrienF.G., MalkowskiM.J., PearsonJ.C., StephensA.J., GiffardP.M., and Australian Group for Antimicrobial resistance. 2004. Genetic diversity among community methicillin-resistant Staphylococcus aureus strains causing outpatient infections in Australia. J. Clin. Microbiol., 42:4735–4743.
23.
Perez-RothE., Lorenzo-DiazF., BatistaN., MorenoA., and Mendez-AlverezS.2004. Tracking methicillin resistant Staphylococcus aureus clones during a 5-year period (1998 to 2002) in a Spanish hospital. J. Clin. Microbiol., 42:4649–4656.
24.
WisplinghoffH., EwertzB., WisplinghoffS., StefanikD., PlumG., Perdreau-RemingtonF., and SeifertH.2005. Molecular evolution of methicillin-resistant Staphylococcus aureus in the metropolitan area of Cologne, Germany, from 1984 to 1998. J. Clin. Microbiol., 43:5445–5451.
25.
McDougalL.K, StewardC.D., KillgoreG.E., ChaitramJ.M., McAllisterS.K., and TenoverF.C.2003. Pulsed-field gel electrophoresis typing of oxacillin resistant Staphylococcus aureus isolates from the United States establishing a national database. J. Clin. Microbiol., 41:5113–5129.
26.
Aires de SousaM., CrisostomoM.I., SanchesI.S., WuJ.S., FuzhongJ., TomaszA., and De lencastreH.2003. Frequent recovery of a single clonal type of multidrug-resistant Staphylococcus aureus from patients in two hospitals in Taiwan and China. J. Clin. Microbiol., 41:159–163.
27.
ArakereG., SavithaN., GöteS., RaginiM., SatishA.K., and DasarathyR.2005. Genotyping of methicillin-resistant Staphylococcus aureus strains from two hospitals in Bangalore, South India. J. Clin. Microbiol., 43:3198–3202.
28.
BudimirA., DeurenbergR.H., PleckoV., VinkC., KalenicS., and StobberinghE.E.2006. Molecular characterization of methicillin-resistant Staphylococcus aureus bloodstream isolates from Croatia. J. Antimicrob. Chem., 57:331–334.
29.
NovickR.P., and E. 2008. Geisinger. Quorum sensing in Staphylococci. Annu. Rev. Genet., 42:541–564.
30.
PainterK.L., KrishnaA., WigneshwerarajS., and EdwardsA.M.2014. What role does the quorum-sensing accessory gene regulator system play during Staphylococcus aureus bacteremia?. Trends Microbiol, 22:676–685.
31.
WrightJ.S., TraberK.E., CorriganR., BensonS.A., MusserJ.M., and NovickR.P.2005. The agr radiation: an early event in the evolution of Staphylococci. J. Bacteriol., 187:5585–5594.
32.
RuudH.D., CornelisV., GuyJ., JaschaO.E., ChristelD., GuyC., JosC., ElsB., SebastianL., HansW., AlexanderW.F., JacquesS., and EllenE.S.. 2005. Different clonal complexes of methicillin-resistant Staphylococcus aureus are disseminated in the Euregio Meuse-Rhine region. Antimicrob. Agents Chem., 49:4263–4271.
33.
WannetW.J.B., HeckM.E.O.C., PluisterG.N., SpalburgE., VanSantenM.G., HuijsdensX.W., TiemersmaE., and deNeelingA.J.. 2004. Panton-Valentine leukocidin positive MRSA in 2003: the Dutch situation. Euro. Surveill., 9:3–4.
34.
TekeliA., OcalD., OzmenB.B., AlayI.M., KarahanZ.C., and DolapciI.2014. Investigation of sasX gene positivity in methicillin-resistant Staphylococcus aureus strains isolated at Ankara University School of Medicine İbni Sina Hospital between years 2002–2013. Abstract presented at 8th National Congress of Molecular and Diagnostic Microbiology, Ankara, June 4–7. Abstract no. OP-006.
35.
Aires de SousaM., and De LencastreH.2003. Evolution of sporadic isolates of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals and their similarities to isolates of community-acquired MRSA. J. Clin. Microbiol., 41:3806–3815.
36.
OztopA.Y., PınarbasiH., KocagözS., BakıcıM.Z., and BakırM.2004. Molecular genotyping of methicillin resistant Staphylococcus aureus strains in a teaching hospital in Turkey. Microb. Drug Resist., 10:154–159.
37.
van BelkumA., van LeeuwenW., VerkooyenR., SacılıkS.C., CokmusC., and VerbrughH.1997. Dissemination of a single clone of methicillin-resistant Staphylococcus aureus among Turkish hospitals. J. Clin. Microbiol, 35:978–981.
38.
ErgonM.C., BicmenM., and GulayZ.2010. Molecular typing of the dominant dominant methicilline resistant Staphylococcus aureus strain at Dokuz Eylul University Hospital. ANKEM Derg., 24:65–70.
