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bla OXA-48 pozitif K. pneumoniae İzolatlarında Fenotipik Karbapenemaz Üretiminin Modifiye Hodge ve Karbapenemaz İnaktivasyon Testleri ile Değerlendirilmesi

Year 2020, Volume: 8 Issue: 1, 1 - 10, 20.04.2020

Reyhan Yiş , E. Deniz Bayram Özlem Yüksel Ergin

https://doi.org/10.37696/nkmj.603254

Abstract

Amaç:
Son
yıllarda özellikle uzun süre hastanede yatan, immun supresyonu olan,  invaziv araç ve geniş spektrumlu antibi­yotik
kullanılan hastalarda kolonizasyon oranları gittikçe artmakta olan K.pneumoniae,
dirençli enfeksiyonlara yol açmaktadır.  Çalışmamızda laboratuvarımıza
gönderilmiş olan klinik örneklerden izole ettiğimiz çoklu dirençli K. pneumoniae izolatlarında karbapenem
direncinin fenotipik ve genotipik yöntemlerle saptanması
amaçlanmıştır.



Yöntemler: Temmuz
2014’te çeşitli servislerden labora­tuvarımıza gelen örneklerden izole edilen,
51 adet K. pneumoniae değerlendirilmiştir. İzolatların tür düzeyinde
tanımlanması ve antibiyotik duyarlılık tesleri otomati­ze sistem ile çalışılmıştır.
Karbapenem direnci gradient test stripleri ile doğrulanmıştır. Karbapenemaz direnç genlerinin
saptanması ‘in-house’ polimeraz zincir tepkimesi (PZT) ile
gerçekleştirilmiştir. Tüm izolatlara fenotipik testlerden Modifiye Hodge
testi (MHT) ve Karbapenemaz İnaktivasyon Testi
(CIM Testi) uygulanmıştır. MHT ve CIM testi uyumsuz sonuç veren izolat için her
iki test tüm karbapenemler ile tekrar çalışılmıştır.



Bulgular: İmipenem (IPM),
meropenem
(MEM) ve ertapenem (ETP) direnç oranları sırasıyla %78,43,
%60,78 ve %96,08 olarak belirlenmiştir. MHT ile 51 adet izolattan 42 ‘sinde,
CIM testi ile 43 adet izolatta pozitiflik saptanmıştır. PCR sonuçlarına göre
izolatların 42 tanesinde blaOXA-48 geni bulunduğu belirlenmiş olup, diğer
karbapenemaz genleri saptanmamıştır. blaOXA-48 geni ve diğer karbapenemeaz
genleri negatif olarak saptanan 9 izolatın tamamında MHT, 8 izolatta ise CIM Testi ile negatiflik saptanmıştır.



Sonuç:
MHT ve CIM, karbapenemaz varlığının saptanmasında basit ve hızla
uygulanabilecek yöntemlerdir. CIM, değerlendirilmesi daha kolay, bir tarama
testi olarak faydalı olabilecek bir yöntem olsa da, yalancı negatif ve yalancı
pozitif sonuçlar açısından dikkatli olmayı gerektirmektedir. Testin diğer
karbapenem disklerinin kullanımı yoluyla modifiye edilmesinin yalancı
negatiflikleri azaltabileceğini düşünüyoruz. 

Keywords

OXA-48 Karbapenem direnci Modifiye Hodge Testi Karbapenem İnaktivasyon Testi PCR

References

  • 1. Aktaş Z, Kayacan CB, Schneider I, Can B, Midilli K, Bauernfeind A. Carbapenem-hydrolyzing oxa¬cillinase, OXA-48, persist in Klebsiella pneumoni¬ae in İstanbul, Turkey, Chemotherapy 2008; 54(2):101-6.
  • 2. Livermore DM. Has the era of untreatable infecti¬ons arrived ? J Antimicrob Chemother 2009;64(Suppl 1):29-36.
  • 3. Aktaş Z, Satana D, Kayacan Ç, Ozbek B, Gurler N, Somer A et al. Carbapenem resistance in Turkey: repeat report on OXA-48 in Klebsiella pneumoniae and first report on IMP-1 beta-lactamase in Escherichia coli, Afr J Microbiol Res 2012;6(17):3874-8.
  • 4. Carrer A, Poirel L, Eraksoy H, Cagatay A, Badur S, Nordmann P. Spread of OXA-48 positive carbapenem-resistant Klebsiella pneumoniae iso¬letes in İstanbul, Turkey, Antimicrob Agents Chemother 2008;52(8):2950-4.
  • 5. Nordmann P, Naas T,Poirel L. Global spread of carbapenemase-producing Enterobacteriaceae, Emerg Infec Dis 2011;17(10):1791-8.
  • 6. Maltezou HC. Metallo-beta-lactamases in Gram negative bacteria:introducing the era of panresis¬tance ? Int J Antimicrob Agents 2009;33(5):405.
  • 7. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing, 21st informational supplement. CLSI Document M100-S21, 2013. CLSI, Wayne, PA.
  • 8. European Committee on Antimicrobial Susceptibility Testing. The EUCAST guideline on detection of resistance mechanisms, v 1.0. Available at: http://www.eucast.org/resistance_mechanisms/
  • 9. Clinical and Laboratory Standards Institute. 2016. Performance Standards for Antimicrobial Susceptibility Testing: 26th Informational Supplement. CLSI M100-S26. CLSI, Wayne, PA
  • 10. van der Zwaluw K, de Haan A, Pluister GN, Bootsma HJ, de Neeling AJ, Schouls LM. The carbapenem inactivation method (CIM), a simple and low-cost alternative for the Carba NP test to assess phenotypic carbapenemase activity in gram-negative rods. PLoS One 2015; 10:e0123690.
  • 11. Nordmann P, Poirel L, Carrër A, Toleman MA, Walsh TR. How to detect NDM-1 producers. J Clin Microbiol. 2011;49(2):718-721.
  • 12. Poirel L, Héritier C, Tolün V, Nordmann P. Emergence of oxacillinase-mediated resistance to IPM in Klebsiella pneumoniae. Antimicrob Agents Chemother. 2004;48(1):15-22.
  • 13. Pitout JD, Gregson DB, Poirel L, McClure JA, Le P, Church DL. Detection of Pseudomonas aeruginosa producing metallo-beta-lactamases in a large centralized laboratory. J Clin Microbiol. 2005;43(7):3129-35.
  • 14. Yigit H, Queenan AM, Anderson GJ, Domenech-Sanchez A, Biddle JW, Steward CD, et al. Novel carbapenem-hydrolyzing beta-lactamase, KPC-1, from a carbapenem-resistant strain of Klebsiella pneumoniae. Antimicrob Agents Chemother 2001;45:1151–1161
  • 15. Bush K, Jacoby GA. Updated functional classification of beta- lactamases. Antimicrob Agents Chemother. 2010;54(3):969–76.
  • 16. Bartolini A, Frasson I, Cavallaro A, Richter SN, Palu G. Comparison of phenotypic methods for the detection of carbapenem non-susceptible Enterobacteriaceae. Gut Pathog 2014; 6:13.
  • 17. Voets GM, Fluit AC, Scharringa J, Cohen Stuart J, Leverstein-van Hall MA. A set of multiplex PCRs for genotypic detection of extended-spectrum b-lactamases, carbapenemases, plasmid-mediated AmpC b-lactamases and OXA b-lactamases. Int J Antimicrob Agents 2011; 37: 356–359.
  • 18. Poirel L, Walsh TR, Cuvillier V, Nordmann P. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis 2011; 70: 119–125.
  • 19. Chen L, Mediavalla JR, Endimiani A, Rosenthal ME, Zhao Y, Bonomo RA et al. Multiplex real-time PCR assay for detection and classification of Klebsiella pneumoniae carbapenemase gene (blaKPC) variants. J Clin Microbiol 2011; 49: 579–585.
  • 20. Monteiro J, Widen RH, Pignatari A, Kubasek C, Silbert S. Rapid detection of carbapenemase genes by multiplex real-time PCR. J Antimicrob Chemother 2012; 67: 906–909.
  • 21. Nordmann P, Poirel L. Strategies for identification of carbapenemase-producing Enterobacteriaceae. J Antimicrob Chemother. 2013;68(3):487-9. doi: 10.1093/jac/dks426. Epub 2012 Oct 26.
  • 22. Cohen Stuart J, Leverstein-Van Hall, Dutch MA. Working Party on the detection of highly resistant microorganisms. Guideline for phenotypic screening and confirmation of carbapenemases in Enterobacteriaceae. Int J Antimicrobial Agents 2010; 36:205-10. https://doi.org/10.1016/j.ijantimicag.2010.05.014
  • 23. Voulgari E, Poulou A, Koumaki V, Tsakris A. Carbapenemase-producing Enterobacteriaceae: now that the storm is finally here, how will timely detection help us fight back? Future Microbiol 2013; 8(1): 27-39.
  • 24. Birgy A, Bidet P, Genel N, Doit C, Decré D, Arlet G, et al. Phenotypic screening of carbapenemases and associated β-lactamases in carbapenem-resistant Enterobacteriaceae. J Clin Microbiol 2012; 50(4): 1295-302.
  • 25. Hrabák J, Chudáčková E, Papagiannitsis CC. Detection of carbapenemases in Enterobacteriaceae: a challenge for diagnostic microbiological laboratories. Clin Microbiol Infect 2014; 20(9): 839-53.
  • 26. Ellis C, Chung C, Tijet N, Patel SN, Desjardins M, Melano RG, et al. OXA-48-like carbapenemase-producing Enterobacteriaceae in Ottawa, Canada. Diagn Microbiol Infect Dis 2013; 76(3): 399-400.
  • 27. Potron A, Poirel L, Rondinaud E, Nordmann P. Intercontinental spread of OXA-48 beta-lactamaseproducing Enterobacteriaceae over a 11-year period, 2001 to 2011. Euro Surveill 2013; 18(31): 20549.
  • 28. Doumith M, Ellington MJ, Livermore DM, Woodford N. Molecular mechanisms disrupting porin expression in ETP-resistant Klebsiella and Enterobacter spp. clinical isolates from the UK. J Antimicrob Chemother. 2009;63:659-67
  • 29. Girlich D, Poirel L, Nordmann P. Value of the modified Hodge test for detection of emerging carbapenemases in Enterobacteriaceae. Clin Microbiol 2012; 50(2): 477-9.
  • 30. Baran I, Aksu N. Phenotypic and genotypic characteristics of carbapenem-resistant Enterobacteriaceae in a tertiary-level reference hospital in Turkey. Ann Clin Microbiol Antimicrob. 2016; 15: 20.
  • 31. Bayramoğlu G, Uluçam G, Gençoğlu Özgür Ç, Kılıç AO, Aydın F. Comparison of the modified Hodge test and the Carba NP test for detection of carbapenemases in Enterobacteriaceae isolates. Mikrobiyol Bul. 2016;50(1):1-10.
  • 32. Yamada K, Kashiwa M, Arai K, Nagano N, Sait R, Comparison of the Modified-Hodge test, Carba NP test, and carbapenem inactivation method as screening methods for carbapenemase-producing Enterobacteriaceae J Microbiol Methods. 2016;128:48-51.
  • 33. Yıldız SS, Kaşkatepe B, Avcıküçük H, Öztürk Ş. Performance of CarbaNP and CIM tests in OXA-48 carbapenemase-producing Enterobacteriaceae. Acta Microbiol Immunol Hung. 2017;64(1):9-16.
  • 34. Bayraktar B, Barış A, Malkoçoğlu G, Erdemir D, Kına N. Comparison of Carba NP-Direct, Carbapenem Inactivation Method, and β-CARBA Tests for Detection of Carbapenemase Production in Enterobacteriaceae. Microb Drug Resist. 2019;25(1):97-102. doi: 10.1089/mdr.2017.0427

Year 2020, Volume: 8 Issue: 1, 1 - 10, 20.04.2020

Reyhan Yiş , E. Deniz Bayram Özlem Yüksel Ergin

https://doi.org/10.37696/nkmj.603254

Abstract

References

  • 1. Aktaş Z, Kayacan CB, Schneider I, Can B, Midilli K, Bauernfeind A. Carbapenem-hydrolyzing oxa¬cillinase, OXA-48, persist in Klebsiella pneumoni¬ae in İstanbul, Turkey, Chemotherapy 2008; 54(2):101-6.
  • 2. Livermore DM. Has the era of untreatable infecti¬ons arrived ? J Antimicrob Chemother 2009;64(Suppl 1):29-36.
  • 3. Aktaş Z, Satana D, Kayacan Ç, Ozbek B, Gurler N, Somer A et al. Carbapenem resistance in Turkey: repeat report on OXA-48 in Klebsiella pneumoniae and first report on IMP-1 beta-lactamase in Escherichia coli, Afr J Microbiol Res 2012;6(17):3874-8.
  • 4. Carrer A, Poirel L, Eraksoy H, Cagatay A, Badur S, Nordmann P. Spread of OXA-48 positive carbapenem-resistant Klebsiella pneumoniae iso¬letes in İstanbul, Turkey, Antimicrob Agents Chemother 2008;52(8):2950-4.
  • 5. Nordmann P, Naas T,Poirel L. Global spread of carbapenemase-producing Enterobacteriaceae, Emerg Infec Dis 2011;17(10):1791-8.
  • 6. Maltezou HC. Metallo-beta-lactamases in Gram negative bacteria:introducing the era of panresis¬tance ? Int J Antimicrob Agents 2009;33(5):405.
  • 7. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing, 21st informational supplement. CLSI Document M100-S21, 2013. CLSI, Wayne, PA.
  • 8. European Committee on Antimicrobial Susceptibility Testing. The EUCAST guideline on detection of resistance mechanisms, v 1.0. Available at: http://www.eucast.org/resistance_mechanisms/
  • 9. Clinical and Laboratory Standards Institute. 2016. Performance Standards for Antimicrobial Susceptibility Testing: 26th Informational Supplement. CLSI M100-S26. CLSI, Wayne, PA
  • 10. van der Zwaluw K, de Haan A, Pluister GN, Bootsma HJ, de Neeling AJ, Schouls LM. The carbapenem inactivation method (CIM), a simple and low-cost alternative for the Carba NP test to assess phenotypic carbapenemase activity in gram-negative rods. PLoS One 2015; 10:e0123690.
  • 11. Nordmann P, Poirel L, Carrër A, Toleman MA, Walsh TR. How to detect NDM-1 producers. J Clin Microbiol. 2011;49(2):718-721.
  • 12. Poirel L, Héritier C, Tolün V, Nordmann P. Emergence of oxacillinase-mediated resistance to IPM in Klebsiella pneumoniae. Antimicrob Agents Chemother. 2004;48(1):15-22.
  • 13. Pitout JD, Gregson DB, Poirel L, McClure JA, Le P, Church DL. Detection of Pseudomonas aeruginosa producing metallo-beta-lactamases in a large centralized laboratory. J Clin Microbiol. 2005;43(7):3129-35.
  • 14. Yigit H, Queenan AM, Anderson GJ, Domenech-Sanchez A, Biddle JW, Steward CD, et al. Novel carbapenem-hydrolyzing beta-lactamase, KPC-1, from a carbapenem-resistant strain of Klebsiella pneumoniae. Antimicrob Agents Chemother 2001;45:1151–1161
  • 15. Bush K, Jacoby GA. Updated functional classification of beta- lactamases. Antimicrob Agents Chemother. 2010;54(3):969–76.
  • 16. Bartolini A, Frasson I, Cavallaro A, Richter SN, Palu G. Comparison of phenotypic methods for the detection of carbapenem non-susceptible Enterobacteriaceae. Gut Pathog 2014; 6:13.
  • 17. Voets GM, Fluit AC, Scharringa J, Cohen Stuart J, Leverstein-van Hall MA. A set of multiplex PCRs for genotypic detection of extended-spectrum b-lactamases, carbapenemases, plasmid-mediated AmpC b-lactamases and OXA b-lactamases. Int J Antimicrob Agents 2011; 37: 356–359.
  • 18. Poirel L, Walsh TR, Cuvillier V, Nordmann P. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis 2011; 70: 119–125.
  • 19. Chen L, Mediavalla JR, Endimiani A, Rosenthal ME, Zhao Y, Bonomo RA et al. Multiplex real-time PCR assay for detection and classification of Klebsiella pneumoniae carbapenemase gene (blaKPC) variants. J Clin Microbiol 2011; 49: 579–585.
  • 20. Monteiro J, Widen RH, Pignatari A, Kubasek C, Silbert S. Rapid detection of carbapenemase genes by multiplex real-time PCR. J Antimicrob Chemother 2012; 67: 906–909.
  • 21. Nordmann P, Poirel L. Strategies for identification of carbapenemase-producing Enterobacteriaceae. J Antimicrob Chemother. 2013;68(3):487-9. doi: 10.1093/jac/dks426. Epub 2012 Oct 26.
  • 22. Cohen Stuart J, Leverstein-Van Hall, Dutch MA. Working Party on the detection of highly resistant microorganisms. Guideline for phenotypic screening and confirmation of carbapenemases in Enterobacteriaceae. Int J Antimicrobial Agents 2010; 36:205-10. https://doi.org/10.1016/j.ijantimicag.2010.05.014
  • 23. Voulgari E, Poulou A, Koumaki V, Tsakris A. Carbapenemase-producing Enterobacteriaceae: now that the storm is finally here, how will timely detection help us fight back? Future Microbiol 2013; 8(1): 27-39.
  • 24. Birgy A, Bidet P, Genel N, Doit C, Decré D, Arlet G, et al. Phenotypic screening of carbapenemases and associated β-lactamases in carbapenem-resistant Enterobacteriaceae. J Clin Microbiol 2012; 50(4): 1295-302.
  • 25. Hrabák J, Chudáčková E, Papagiannitsis CC. Detection of carbapenemases in Enterobacteriaceae: a challenge for diagnostic microbiological laboratories. Clin Microbiol Infect 2014; 20(9): 839-53.
  • 26. Ellis C, Chung C, Tijet N, Patel SN, Desjardins M, Melano RG, et al. OXA-48-like carbapenemase-producing Enterobacteriaceae in Ottawa, Canada. Diagn Microbiol Infect Dis 2013; 76(3): 399-400.
  • 27. Potron A, Poirel L, Rondinaud E, Nordmann P. Intercontinental spread of OXA-48 beta-lactamaseproducing Enterobacteriaceae over a 11-year period, 2001 to 2011. Euro Surveill 2013; 18(31): 20549.
  • 28. Doumith M, Ellington MJ, Livermore DM, Woodford N. Molecular mechanisms disrupting porin expression in ETP-resistant Klebsiella and Enterobacter spp. clinical isolates from the UK. J Antimicrob Chemother. 2009;63:659-67
  • 29. Girlich D, Poirel L, Nordmann P. Value of the modified Hodge test for detection of emerging carbapenemases in Enterobacteriaceae. Clin Microbiol 2012; 50(2): 477-9.
  • 30. Baran I, Aksu N. Phenotypic and genotypic characteristics of carbapenem-resistant Enterobacteriaceae in a tertiary-level reference hospital in Turkey. Ann Clin Microbiol Antimicrob. 2016; 15: 20.
  • 31. Bayramoğlu G, Uluçam G, Gençoğlu Özgür Ç, Kılıç AO, Aydın F. Comparison of the modified Hodge test and the Carba NP test for detection of carbapenemases in Enterobacteriaceae isolates. Mikrobiyol Bul. 2016;50(1):1-10.
  • 32. Yamada K, Kashiwa M, Arai K, Nagano N, Sait R, Comparison of the Modified-Hodge test, Carba NP test, and carbapenem inactivation method as screening methods for carbapenemase-producing Enterobacteriaceae J Microbiol Methods. 2016;128:48-51.
  • 33. Yıldız SS, Kaşkatepe B, Avcıküçük H, Öztürk Ş. Performance of CarbaNP and CIM tests in OXA-48 carbapenemase-producing Enterobacteriaceae. Acta Microbiol Immunol Hung. 2017;64(1):9-16.
  • 34. Bayraktar B, Barış A, Malkoçoğlu G, Erdemir D, Kına N. Comparison of Carba NP-Direct, Carbapenem Inactivation Method, and β-CARBA Tests for Detection of Carbapenemase Production in Enterobacteriaceae. Microb Drug Resist. 2019;25(1):97-102. doi: 10.1089/mdr.2017.0427
There are 34 citations in total.

Details

Primary Language Turkish
Subjects Clinical Sciences
Journal Section Orginal Article
Authors

Reyhan Yiş 0000-0001-5774-0315

E. Deniz Bayram This is me 0000-0003-2299-6331

Özlem Yüksel Ergin This is me 0000-0002-5392-9345

Publication Date April 20, 2020
Published in Issue Year 2020 Volume: 8 Issue: 1

Cite

APA Yiş, R., Bayram, E. D., & Yüksel Ergin, Ö. (2020). bla OXA-48 pozitif K. pneumoniae İzolatlarında Fenotipik Karbapenemaz Üretiminin Modifiye Hodge ve Karbapenemaz İnaktivasyon Testleri ile Değerlendirilmesi. Namık Kemal Tıp Dergisi, 8(1), 1-10. https://doi.org/10.37696/nkmj.603254
AMA Yiş R, Bayram ED, Yüksel Ergin Ö. bla OXA-48 pozitif K. pneumoniae İzolatlarında Fenotipik Karbapenemaz Üretiminin Modifiye Hodge ve Karbapenemaz İnaktivasyon Testleri ile Değerlendirilmesi. NKMJ. April 2020;8(1):1-10. doi:10.37696/nkmj.603254
Chicago Yiş, Reyhan, E. Deniz Bayram, and Özlem Yüksel Ergin. “Bla OXA-48 Pozitif K. Pneumoniae İzolatlarında Fenotipik Karbapenemaz Üretiminin Modifiye Hodge Ve Karbapenemaz İnaktivasyon Testleri Ile Değerlendirilmesi”. Namık Kemal Tıp Dergisi 8, no. 1 (April 2020): 1-10. https://doi.org/10.37696/nkmj.603254.
EndNote Yiş R, Bayram ED, Yüksel Ergin Ö (April 1, 2020) bla OXA-48 pozitif K. pneumoniae İzolatlarında Fenotipik Karbapenemaz Üretiminin Modifiye Hodge ve Karbapenemaz İnaktivasyon Testleri ile Değerlendirilmesi. Namık Kemal Tıp Dergisi 8 1 1–10.
IEEE R. Yiş, E. D. Bayram, and Ö. Yüksel Ergin, “bla OXA-48 pozitif K. pneumoniae İzolatlarında Fenotipik Karbapenemaz Üretiminin Modifiye Hodge ve Karbapenemaz İnaktivasyon Testleri ile Değerlendirilmesi”, NKMJ, vol. 8, no. 1, pp. 1–10, 2020, doi: 10.37696/nkmj.603254.
ISNAD Yiş, Reyhan et al. “Bla OXA-48 Pozitif K. Pneumoniae İzolatlarında Fenotipik Karbapenemaz Üretiminin Modifiye Hodge Ve Karbapenemaz İnaktivasyon Testleri Ile Değerlendirilmesi”. Namık Kemal Tıp Dergisi 8/1 (April 2020), 1-10. https://doi.org/10.37696/nkmj.603254.
JAMA Yiş R, Bayram ED, Yüksel Ergin Ö. bla OXA-48 pozitif K. pneumoniae İzolatlarında Fenotipik Karbapenemaz Üretiminin Modifiye Hodge ve Karbapenemaz İnaktivasyon Testleri ile Değerlendirilmesi. NKMJ. 2020;8:1–10.
MLA Yiş, Reyhan et al. “Bla OXA-48 Pozitif K. Pneumoniae İzolatlarında Fenotipik Karbapenemaz Üretiminin Modifiye Hodge Ve Karbapenemaz İnaktivasyon Testleri Ile Değerlendirilmesi”. Namık Kemal Tıp Dergisi, vol. 8, no. 1, 2020, pp. 1-10, doi:10.37696/nkmj.603254.
Vancouver Yiş R, Bayram ED, Yüksel Ergin Ö. bla OXA-48 pozitif K. pneumoniae İzolatlarında Fenotipik Karbapenemaz Üretiminin Modifiye Hodge ve Karbapenemaz İnaktivasyon Testleri ile Değerlendirilmesi. NKMJ. 2020;8(1):1-10.