Ankara’daki çeşitli hastanelerden elde edilen Acinetobacter baumannii izolatları arasındaki klonal ilişkinin gösterilmesi ve antibiyotik dirençlerinin belirlenmesi

Year 2016, Volume: 73 Issue: 3, 199 - 210, 01.09.2016

Hünkar Şahin Ufuk Önde Ali Kudret Adiloğlu Ayşe Esra Karakoç Cemal Bulut Ziya Cibali Açıkgöz Gül Bahar Erdem Gülşen Hasçelik Ayşegül Öztürk Coşkun

Abstract

Amaç: Acinetobacter baumannii, özellikle yoğun bakım ünitelerinde mortalite ve morbiditeyi arttıran hastane kaynaklı enfeksiyonların en önemli nedenlerinden biridir. Bu bakteri, pnömoni, bakteriyemi, idrar yolu enfeksiyonu, yara enfeksiyonu ve menenjit gibi çeşitli enfeksiyonlardan izole edilebilir. Bu çalışmanın amacı Ankara’daki çeşitli hastanelerden Kasım 2009 ile Aralık 2011 tarihleri arasında elde edilen 99 A. baumannii izolatı arasındaki klonal ilişkiyi göstermek ve antibiyotik dirençlerini belirlemektir. Yöntem: Acinetobacter baumannii izolatlarının tanımlanması ve antibiyotik paternleri VITEK 2 bioMérieux, France sistemi ile yapıldı. Çalışmada antimikrobiyal ajan olarak amikasin, siprofloksasin, tetrasiklin, netilmisin, sulbaktam/ ampisilin, trimetoprim/sulfametoksazol, seftazidim, gentamisin, levofloksasin, meropenem, imipenem, piperasilin, piperasilin/tazobaktam, sefoperazon/ sulbaktam, sefepim, tigesiklin ve kolistin kullanıldı. Acinetobacter izolatlarının klonal ilişkisi Rep-PCR yöntemi ile araştırıldı. Bulgular: Çalışmaya dâhil edilen 99 A. baumannii izolatının %99’u imipeneme ve meropeneme dirençli bulundu. Kolistine dirençli suş saptanmadı %0 . Piperasilin ve piperasilin/tazobaktama tüm suşların dirençli olduğu gözlendi %100 . Siprofloksasine %99, ampisilin/sulbaktama %98, seftazidime %97, sefepime %96, levofloksasine %85, tetrasikline %80, trimetoprim/sulfametoksazole %71, amikasine %59, gentamisin ve netilmisine %56 direnç görüldü. Tigesiklin için dirençliler %5, orta duyarlılar %27 ve duyarlılar ise %68 olarak saptandı. Acinetobacter baumannii izolatlarının dört patern oluşturduğu görüldü. İlk 3 paternin bir grup oluşturduğu ve dördüncü paternden farklı olduğu görüldü. Birinci patern P1 ’de 14 suş, ikinci patern P2 ’de 37 suş, üçüncü patern P3 ’de 36 suş bulunmaktaydı. Dördüncü patern P4 ’de ise 12 suş vardı. İlk üç paternin oluşturduğu grup ile dördüncü paternin farklı oldukları ve aralarında üçten fazla bant farklılığının olduğu görüldü. İlk üç paternin ise aralarındaki bir bant farklılığı ile benzer oldukları görüldü. Ayrıca, her dört paternin kendi içinde bant farkı göstermeksizin aynı oldukları saptandı. Her dört paternde de farklı hastanelerden suşların bulunduğu saptandı. Böylece her dört hastaneye muhtemelen hasta transferleriyle aynı Acinetobacter baumannii suşlarının yayılmış olduğu gösterildi. Sonuç: Bu hastanelerde, çalışmanın devamı olarak A. baumannii’nin direnç mekanizmaları ve direnç genleriyle ilgili araştırma yapılabilir. Ayrıca, hastalara ilişkin risk faktörleri belirlenerek karbapenem ve diğer antibiyotiklerin dirençlerinden sorumlu izolatlara ilişkin epidemiyolojik veriler daha kapsamlı bir şekilde ortaya konulabilir.

Keywords

Acinetobacter baumannii, klonal ilişki, rep-PCR, antibiyotik direnci

Evaluation Of Clonal Relationship Between Acinetobacter baumannii Isolates and Determination Of Resistance To Antibiotics Which Were Obtained from Different Hospitals In Ankara

Abstract

Objective: Acinetobacter baumannii is one of the most important cause of hospital acquired infections which raises the rates of mortality and morbidity especially in intensive care units. Acinetobacter baumannii may be isolated from various infections such as pneumonia, bacteremia, urinary track infections, wound infections and meningitis.The aim of this study is to determine the clonal relationship and determination of antibiotic resistances between 99 Acinetobacter baumannii isolates which were obtained from different hospitals in Ankara between November 2009 and December 2011. Method: Identification of A. baumannii isolates and antibiotic susceptibilities were performed by Vitek 2 bioMérieux, France system. In this study, amikacin, ciprofloxacin, tetracyclin, netilmicin, sulbactam/ampicillin, trimethoprim/sulfamethoxazole, ceftazidime, gentamicin, levofloxacin, meropenem, imipenem, piperacillin, piperacillin/tazobactam, cefoperazone/sulbactam, cefepime, tigecycline, and colistin were used as antimicrobial agents. The clonal relationship of Acinetobacter isolates was analysed by Rep-PCR method. Results: Ninety nine percent of A. baumannii isolates was resistant to imipenem and meropenem. There was no resistant strain to colistine 0% . All of the strains were resistant to piperacillin and piperacillin/tazobactam 100% . Ciprofloxacin was 99%, sulbactam/ampicillin 98%, ceftazidime was 97%, cefepime was 96%, levofloxacin was 85%, tetracycline was 80%, trimethoprim/sulfamethoxazole was 71% resistant. Resistance to amikacin was 59% and resistance to gentamicin and netilmicin were both 56%. Five percent of strains were resistant, 27% were intermediate susceptile and 68% were susceptile to tigecycline. There were four patterns. P1, P2 and P3 gathered to one group G1 and they were similar. P4 was different from them. P1 had 14, P2 had 37, P3 had 36 and P4 had 12 strains. There were more than three bands difference between G1 and P4 which showed that they were different strains. Band difference between G1 P1, P2, P3 was only one and this showed us that they were similar. In the other hand, there were not any band diffirence in P1, P2, P3 and P4. All of the patterns were consisted of strains from different hospitals.We determined that the same strains spread to four different hospitals probably by patient transfers. Conclusion: A study can be organized to determine resistance genes and resistance mechanisms of A. baumannii in these hospitals as continuation of this study. Moreover, a comprehensive epidemiologic data can be revealed by determining the risk factors of patients relevant to those responsible isolates for carbapenem and other antibiotics resistance.

Keywords

Acinetobacter baumannii, clonal relationship, rep-PCR, antibiotic resistance

References

• 1. Koneman EW, Allen SD, Janda WM, Schreckenberger PC, Winn WC, Woods G. Koneman’s Color Atlas and Textbook of Diagnostic Microbiology; 6’th ed. Lippincott Philadelphia. 2006:316–355.
• 2. Bergogne-Berezin E, Towner KJ. Acinetobacter spp. as nosocomial pathogens: microbiological, clinical, and epidemilogical features. Clin Microbiol Rev, 1996;9:148–65.
• 3. Villegas MV, Hartstein AI. Acinetobacter outbreaks, 1977-2000. Infection Control and Hospital Epidemiology 2003;24(4):284-95.
• 4. Beck-Sague CM, Jarvis WR, Brook JH, et al. Epidemic bacteremia due to Acinetobacter baumannii in five intensive care unit. Am J Epidemiol 1990;132:723–33.
• 5. Lortholary O, Fagon J-Y, Hoi AB, et al. Nosocomial acquisition of multiresistant Acinetobacter baumannii: risk factors and prognosis. Clin Infect Dis, 1995;20:790–96.
• 6. Jain R, Danziger LH. Multidrug-resistant Acinetobacter infections: an emerging challenge to clinicians. Ann Pharmacother, 2004;38:1449–59.
• 7. Li J, Nation RL, Milne RW, Turnidge JD, Coulthard K. Evaluation of colistin as an agent against multiresistant gram-negative bacteria. Int J Antimicrob Agents 2005;25:11–25.
• 8. Webster CA, Crove M, Humphreys H, Towner KJ. Surveillance of and adult intensive care unit for long-term persistence of a multi-resistant strain of Acinetobacter baumannii. J Clin Microbiol, 1998;17:171–76.
• 9. Corbella X, Pujol M, Ayast J, Sendra M, Ardanury C, Dominguez MA, et al. Relevance of digestive tract colonization in the epidemiology of nasocomial infections due to multi resistant Acinetobacter baumannii. Clin Microbiol Infect Dis, 1996;23:329– 34.
• 10. D’Agata EMC, Thayer V, Schaffner W. An outbreak of Acinetobacter baumannii: The importance of cross-transmission. Infection Control and Hospital Epidemiology 2000; 21: 588-591.
• 11. Bayat A, Shaaban H, Dodgsonb A, Dunna KW. Implications for burns unit design following outbreak of multi-resistant Acinetobacter infection in ICU and Burns Unit. Burns 2003; 29:303–06.
• 12. Husni R N, Goldstein L S, Arroliga A C, Hall G S, Fatica C, Stoller J K, Gordon S M. Risk factors for an outbreak of multi-drug-resistant acinetobacter nosocomial pneumonia among intubated patients, Chest 1999;115;1378-82
• 13. Munoz-Price LS, Weinstein RA. Acinetobacter infection. N Engl J Med. 2008 Mar 20;358(12):1271- 81.
• 14. Saeed S, Fakih MG, Riederer K, et al. Interinstitutional and intrainstitutional transmission of a strain of Acinetobacter baumannii detected by molecular analysis: Comparison of pulsed-field gel electrophoresis and repetitive sequence-based polymerase chain reaction. Infect Control Hosp Epidemiol, 2006;27:981-3.
• 15. Bou G, Cervero G, Dominguez MA, Quereda C, Martinez-Beltran J. PCR based DNA fingerprinting (REP-PCR, AP-PCR) and pulsed-field gel electrophoresis charecterization of a nosocomial outbreak caused by imipenem and meropenemresistant Acinetobacter baumannii. Clinical Microbiology and Infection 2000; 6:635-43.
• 16. Hsueh PR, LJ, Chen CY, et al. Pandrug-resistant Acinetobacter baumannii causing nosocomial infections in a university hospital, Taiwan. Emerg Infect Dis 2002;8:827–32.
• 17. Erben N, Kiremitçi A, Özgüneş İ. Klinik örneklerden izole edilen Acinetobacter türlerinde genişletilmiş spektrumlu beta-laktamaz ve indüklenebilir beta-laktamaz sıklığının ve antimikrobiyal duyarlılığın değerlendirmesi, Osmangazi Tıp Derg, 2006;28(3):135-46.
• 18. Cisneros JM, Rodríguez-Barío J. Nosocomial bacteremia due to Acinetobacter baumannii: epidemiology, clinical features and treatment, Clinical Microbiology and Infection, 2002; 8(11):687-93.
• 19. Smith PW, Massanari RM. Room humidifiers as the source of Acinetobacter infections. Journal of the American Medical Association, 1997; 237(8):795- 97.
• 20. Villers D, Espaze E, Coste-Byrel M. Nosocomial Acinetobacter baumannii infections: microbiological and clinical epidemiology. Annals of Internal Medicin, 1998;129(3):182-89.
• 21. Balcı M, Bitirgen M, Kandemir B, Turk Arıbaş E, Erayman İ. Nozokomiyal Acinetobacter baumannii suşlarının antibiyotik duyarlılığı. Ankem Dergisi 2010; 24: 28-33.
• 22. Weinbren MJ, Johnson AP, Kaufmann ME, Livermore DM. Acinetobacter spp. isolates with reduced susceptibilities to carbapenems in a UK burns unit. The Journal of antimicrobial chemotherapy. 1998, 41(5):574-6.
• 23. Gözütok F, Çelik İ, Berk E, Aydın B, Güzel D. Hastane infeksiyonu etkeni Acinetobacter baumannii suşlarının antimikrobiyal direnç oranlarının araştırılması. Ankem Derg. 2013;27(1):7-12.
• 24. Souli M, Galani I, Giamarellou H. Emergence of extensively drug- resistant and pandrug-resistant Gram-negative bacilli in Europe. Euro surveillance, 2008;13(47):1-11.
• 25. Turner P, Greenhalgh J. The activity of meropenem and comparators against Acinetobacter strains isolated from European hospitals, 1997- 2000. Clin Microbiol Infect, 2003;9(6):563-67.
• 26. Turner PJ. Meropenem activity against European isolates: report on the MYSTIC (Meropenem Yearly Susceptibility Test Information Collection) 2006 results. Diagn Microbiol Infect Dis, 2008;60(2):185- 92.
• 27. Gür D, Hascelik G, Aydin N, et al. Antimicrobial resistance in Gram-negative hospital isolates: results of the Turkish HITIT-2 surveillance study of 2007. J Chemother, 2009;21(4):383-89.
• 28. Gür D, Gülay Z, Akan ÖA, et al. Türkiye’de hastane izolatı Gram-negatif bakterilerde yeni betalaktam antibiyotiklere direnç ve GSBL tipleri: Çok merkezli HİTİT sürveyansının sonuçları. Mikrobiyol Bul, 2008;42(4):537-44.
• 29. Maragakis L, Perl T. Acinetobacter baumannii: epidemiology, antimicrobial resistance, and treatment options. Clin Infect Dis, 2008;46(8):1254- 63.
• 30. Reboli AC, Houston ED, Monteforte JS, et al. Discrimination of epidemic and sporadic isolates of Acinetobacter baumannii by repetitive element PCR-mediated DNA fingerprinting. J Clin Microbiol 1994; 32(11): 2635-40.