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Investigation of the presence of OXA 48 and mcr-1 genes in carbapenem-resistant Acinetobacter baumannii, Klebsiella pneumoniae and Pseudomonas aeruginosa strains by in house-PCR method

Year 2021, , 118 - 123, 21.12.2021
https://doi.org/10.47582/jompac.1028959

Abstract

Karbapenem dirençli Enterobacteriaceae (KRE)’nin neden olduğu enfeksiyonlar, küresel sağlık için ciddi bir sorundur. Karbapenemler, kapsamlı uygulamaları nedeniyle günümüzde direnç mekanizmaları gelişmiştir. Karbapenemaz sentezleyen izolatların saptanması, dağılımlarının takip edilmesi ve uygun tedavinin sağlanabilmesi açısından oldukça önemlidir. Günümüz ilaç direncinin saptanmasında Real-time-polimeraz zincir reaksiyonu (RT-PZR), başta olmak üzere çeşitli ticari moleküler tanı yöntemleri kullanılmaktadır. Ancak testlerin yapılabilmesi için pahalı ekipmanlara ihtiyaç duyulması nedeni ile, rutin bakteriyoloji laboratuvarlarında uygulanabilecek güvenilir, pratik ve düşük maliyetli testlere ihtiyaç duyulmaktadır.
Bu çalışma, Etlik İhtisas Eğitim ve Araştırma Hastanesi’nde çeşitli servislerde yatan hastalardan izole edilen genişlemiş spektrumlu beta-laktamaz (GSBL) pozitif, karbapenemlere dirençli dirençli, kolistine duyarlı toplam 60 Gram-negatif bakteri (39 suş Acinetobacter baumannii (A. baumannii), 11 suş Klebsiella pneumoniae (K. pneumoniae ve 10 suş Pseudomonas aeruginosa (P. aeruginosa))’de izolatın mcr-1 ve blaOXA-48 gen bölgelerinin varlığını belirlemek amacıyla yapılmıştır. Klinik örnekler konvansiyonel yöntemler ile izole edildikten sonra kesin tanımlama ve antibiyotik duyarlılık testleri otomatize sistemle çalışılmıştır. GSBL ve kolistin direncinin doğrulanması disk difüzyon testi ve gradyen testleri ile yapıldı. PZR sonuçlarına göre A. baumannii suşunun 10 tanesinde, K. pneumoniae suşunun bir tanesinde ve P. aeruginosa suşunun bir tanesinde mcr-1 geni saptandı. A. baumannii suşunun 5 tanesinde, K. pneumoniae suşunun 2 tanesinde blaOXA-48 geni saptandı. A. baumannii suşunun bir tanesinde her iki genin varlığı tespit edilmiştir. Sonuç olarak, karbapenemaz enzimi üretimi ve gen bölgesinin belirlenmesinde hızlı, tekrarlanabilir ve yüksek doğrulukta yeni tanı yöntemlerine ihtiyaç duyulmaktadır.

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References

  • Nukui Y, Ayibieke A, Taniguchi M, et al. Whole-genome analysis of EC129, an NDM-5-, CTX-M-14-, OXA-10- and MCR-1-co-producing Escherichia coli ST167 strain isolated from Japan. J Global Antimicrob Resist 2019; 18: 148-50.
  • Suay-García B, Pérez-Gracia MT. Present and future of carbapenem-resistant Enterobacteriaceae (CRE) infections. Antibiotics (Basel) 2019; 8: 122.
  • Hameed MF, Chen Y, Wang Y, et al. Epidemiological characterization of colistin and carbapenem resistant Enterobacteriaceae in a tertiary: A Hospital from Anhui Province. Infect Drug Resist 2021; 14: 1325-33.
  • Beyrouthy R, Robin F, Lessene A, et al. MCR-1 and OXA-48 In vivo acquisition in KPC-producing Escherichia coli after colistin treatment. Antimicrob Agents Chemother 2017; 61: 8.
  • Di Tella D, Tamburro M, Guerrizio G, Fanelli I, Sammarco ML, Ripabelli G. Molecular epidemiological insights into colistin-resistant and carbapenemases-producing clinical Klebsiella pneumoniaeisolates. Infect Drug Resist 2019; 12: 3783-95.
  • Lee YL, Lu MC, Shao PL, et al. Nationwide surveillance of antimicrobial resistance among clinically important Gram-negative bacteria, with an emphasis on carbapenems and colistin: Results from the Surveillance of Multicenter Antimicrobial Resistance in Taiwan (SMART) in 2018. Int J Antimicrob Agents 2019; 54: 318-28.
  • Demir N. Gram-negatif bakterilerde genişlemiş spektrumlu beta-laktamaz (GSBL) üretimine katkıda bulunan çeşitli risk faktörlerinin araştırılması.İstanbul: Dr Lütfi Kırdar Kartal Eğitim ve Araştırma Hastanesi İnfeksiyon Hastalıkları ve Klinik Mikrobiyoloji Kliniği, Uzmanlik tezi 2006.
  • Hall LM, Livermore DM, Gur D, Akova M, Akalin HE. OXA-11, an extended-spectrum variant of OXA-10 (PSE-2) beta-lactamase from Pseudomonas aeruginosa. Antimicrob Agents Chemother 1993; 37: 1637-44.
  • Codjoe FS, Donkor ES. Carbapenem resistance: a review. Med Sci 2018; 6: 1.
  • Meletis G. Carbapenem resistance: overview of the problem and future perspectives. Therapeutic Advances Infect Dis 2016; 3: 15-21.
  • Nordmann P, Poirel L. The difficult-to-control spread of carbapenemase producers among Enterobacteriaceae worldwide. Clin Microbiol Infect 2014; 20: 821-30.
  • Li Z, Cao Y, Yi L, Liu J-H, Yang Q, editors. Emergent polymyxin resistance: end of an era? Open forum infectious diseases; 2019: Oxford University Press US.
  • Özger HS, Karaşahin Ö, Telli G, Dizbay M. Nozokomiyal Klebsiella türleri arasında karbapenem direnç sıklığı ve fenotipik yöntemlerle direncin değerlendirilmesi. Flora Derg 2012; 17: 103-10.
  • Gharaibeh MH, Shatnawi SQ. An overview of colistin resistance, mobilized colistin resistance genes dissemination, global responses, and the alternatives to colistin: a review. Veterinary World 2019; 12: 1735.
  • Baron S, Hadjadj L, Rolain J-M, Olaitan AO. Molecular mechanisms of polymyxin resistance: knowns and unknowns. Int J Antimicrob Agents 2016; 48: 583-91.
  • Lu H, Zhang X, Zhang Y, et al. First report of multidrug-resistant Escherichia coli isolates co-harbouring mcr-1 and bla(OXA-48) from clinical patients in China. J Global Antimicrob Resist 2018; 13: 945.
  • Lopes E, Saavedra MJ, Costa E, de Lencastre H, Poirel L, Aires-de-Sousa M. Epidemiology of carbapenemase-producing Klebsiella pneumoniae in northern Portugal: Predominance of KPC-2 and OXA-48. J Global Antimicrob Resist 2020; 22: 349-53.
  • Poirel L, Bonnin RA, Nordmann P. Genetic features of the widespread plasmid coding for the carbapenemase OXA-48. Antimicrob Agents Chemother 2012; 56: 559-62.
  • Süzük Yıldız S, Şimşek H, Bakkaloğlu Z, et al. [The Epidemiology of Carbapenemases in Escherichia coli and Klebsiella pneumoniae Isolated in 2019 in Turkey]. Mikrobiyol Bul 2021; 55: 1-16.
  • Çiğdem A, Tuba D, Tuğcan B, Neslihan G, Rıza D. Investigation of carbapenemase and mcr-1 genes in carbapenem-resistant Klebsiella pneumoniae isolates. J Infect Develop Countr 2019; 13: 6.
  • Bilal H, Rehman TU, Khan MA, et al. Molecular Epidemiology of mcr-1, bla (KPC-2,) and bla (NDM-1) Harboring Clinically Isolated Escherichia coli from Pakistan. Infect Drug Resist 2021; 14: 1467-79.
  • Parlak AU, Güdücüoğlu H, Parlak M, Bayram Y, Otlu B. Klebsiella pneumoniaesuşlarında OXA-48 ve alt türevlerinin araştırılması ve fenotipik yansıma. ANKEM Derg 2021; 35: 1-8.
  • Caspar Y, Maillet M, Pavese P, et al. mcr-1 colistin resistance in ESBL-producing Klebsiella pneumoniae, France. Emerg Infect Dis 2017; 23: 874.
  • Robin F, Beyrouthy R, Colot J, et al. MCR-1 in ESBL-producing Escherichia coli responsible for human infections in New Caledonia. J Antimicrob Chemother 2017; 72: 946-7.
  • Gröndahl‐Yli‐Hannuksela K, Lönnqvist E, Kallonen T, et al. The first human report of mobile colistin resistance gene, mcr‐1, in Finland. Apmis 2018; 126: 413-7.
  • Abd El-Baky RM, Masoud SM, Mohamed DS, et al. Prevalence and some possible mechanisms of colistin resistance among multidrug-resistant and extensively drug-resistant Pseudomonas aeruginosa. Infect Drug Resist 2020; 13: 323.
  • Hameed F, Khan MA, Muhammad H, Sarwar T, Bilal H, Rehman TU. Plasmid-mediated mcr-1 gene in Acinetobacter baumannii and Pseudomonas aeruginosa: first report from Pakistan. Revista da Sociedade Brasileira de Medicina Tropical 2019; 52.

Karbapeneme dirençli Acinetobacter baumannii, Klebsiella pneumoniae ve Pseudomanas aeruginosa suşlarında OXA 48 ve mcr-1 gen varlığının in house-PCR yöntemi ile araştırılması

Year 2021, , 118 - 123, 21.12.2021
https://doi.org/10.47582/jompac.1028959

Abstract

Infections caused by carbapenemresistant Enterobacteriaceae (CRE) are a serious problem for global health. Carbapenems have developed resistance mechanisms due to their extensive applications. Identification of isolates expressing carbapenemase is important for that terms of monitoring their distribution and providing appropriate treatment. For detection of drugresistance, it is used various commercial molecular diagnostic methods, especially Real Time-polymerase chain reaction (RT-PCR). However, nowadays due to the needed for expensive equipment to carry out the tests, it is needed reliable, practical and low-cost tests that can be applied in routine bacteriology laboratories.
Our study was carried out to determine the presence of mcr-1 and blaOXA-48 gene regions of 60 ESBL positive, carbapenems resistant, colistin sensitive Gram negative bacteriae (39 strains of Acinetobacter baumannii (A. baumannii), 11 strains of Klebsiella pneumoniae (K. pneumoniae), and 10 strains of Pseudomonas aeruginosa (P. aeruginosa)) from patients hospitalized in various wards of Etlik İhtisas Training and Research Hospital. The clinical samples were isolated by conventional methods, after that precise identificationand antibiotic susceptibility tests were performed with an automated system. Confirmation of ESBL and colistin resistance was done by disc diffusion and gradient testing. According to PCR results, mcr-1 gene was detected in ten A. baumannii strains, one K. pneumoniae and one P. aeruginosa strain. The blaOXA-48 gene was detected in five A. baumannii strains and two K. pneumoniae strains. Both genes were detected in one of the A. baumannii strain. gene. As a result, it was needed new diagnostic methods that rapid, reproducible, highly accurate for determine carbapenemase enzyme production and gene presence.

Project Number

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References

  • Nukui Y, Ayibieke A, Taniguchi M, et al. Whole-genome analysis of EC129, an NDM-5-, CTX-M-14-, OXA-10- and MCR-1-co-producing Escherichia coli ST167 strain isolated from Japan. J Global Antimicrob Resist 2019; 18: 148-50.
  • Suay-García B, Pérez-Gracia MT. Present and future of carbapenem-resistant Enterobacteriaceae (CRE) infections. Antibiotics (Basel) 2019; 8: 122.
  • Hameed MF, Chen Y, Wang Y, et al. Epidemiological characterization of colistin and carbapenem resistant Enterobacteriaceae in a tertiary: A Hospital from Anhui Province. Infect Drug Resist 2021; 14: 1325-33.
  • Beyrouthy R, Robin F, Lessene A, et al. MCR-1 and OXA-48 In vivo acquisition in KPC-producing Escherichia coli after colistin treatment. Antimicrob Agents Chemother 2017; 61: 8.
  • Di Tella D, Tamburro M, Guerrizio G, Fanelli I, Sammarco ML, Ripabelli G. Molecular epidemiological insights into colistin-resistant and carbapenemases-producing clinical Klebsiella pneumoniaeisolates. Infect Drug Resist 2019; 12: 3783-95.
  • Lee YL, Lu MC, Shao PL, et al. Nationwide surveillance of antimicrobial resistance among clinically important Gram-negative bacteria, with an emphasis on carbapenems and colistin: Results from the Surveillance of Multicenter Antimicrobial Resistance in Taiwan (SMART) in 2018. Int J Antimicrob Agents 2019; 54: 318-28.
  • Demir N. Gram-negatif bakterilerde genişlemiş spektrumlu beta-laktamaz (GSBL) üretimine katkıda bulunan çeşitli risk faktörlerinin araştırılması.İstanbul: Dr Lütfi Kırdar Kartal Eğitim ve Araştırma Hastanesi İnfeksiyon Hastalıkları ve Klinik Mikrobiyoloji Kliniği, Uzmanlik tezi 2006.
  • Hall LM, Livermore DM, Gur D, Akova M, Akalin HE. OXA-11, an extended-spectrum variant of OXA-10 (PSE-2) beta-lactamase from Pseudomonas aeruginosa. Antimicrob Agents Chemother 1993; 37: 1637-44.
  • Codjoe FS, Donkor ES. Carbapenem resistance: a review. Med Sci 2018; 6: 1.
  • Meletis G. Carbapenem resistance: overview of the problem and future perspectives. Therapeutic Advances Infect Dis 2016; 3: 15-21.
  • Nordmann P, Poirel L. The difficult-to-control spread of carbapenemase producers among Enterobacteriaceae worldwide. Clin Microbiol Infect 2014; 20: 821-30.
  • Li Z, Cao Y, Yi L, Liu J-H, Yang Q, editors. Emergent polymyxin resistance: end of an era? Open forum infectious diseases; 2019: Oxford University Press US.
  • Özger HS, Karaşahin Ö, Telli G, Dizbay M. Nozokomiyal Klebsiella türleri arasında karbapenem direnç sıklığı ve fenotipik yöntemlerle direncin değerlendirilmesi. Flora Derg 2012; 17: 103-10.
  • Gharaibeh MH, Shatnawi SQ. An overview of colistin resistance, mobilized colistin resistance genes dissemination, global responses, and the alternatives to colistin: a review. Veterinary World 2019; 12: 1735.
  • Baron S, Hadjadj L, Rolain J-M, Olaitan AO. Molecular mechanisms of polymyxin resistance: knowns and unknowns. Int J Antimicrob Agents 2016; 48: 583-91.
  • Lu H, Zhang X, Zhang Y, et al. First report of multidrug-resistant Escherichia coli isolates co-harbouring mcr-1 and bla(OXA-48) from clinical patients in China. J Global Antimicrob Resist 2018; 13: 945.
  • Lopes E, Saavedra MJ, Costa E, de Lencastre H, Poirel L, Aires-de-Sousa M. Epidemiology of carbapenemase-producing Klebsiella pneumoniae in northern Portugal: Predominance of KPC-2 and OXA-48. J Global Antimicrob Resist 2020; 22: 349-53.
  • Poirel L, Bonnin RA, Nordmann P. Genetic features of the widespread plasmid coding for the carbapenemase OXA-48. Antimicrob Agents Chemother 2012; 56: 559-62.
  • Süzük Yıldız S, Şimşek H, Bakkaloğlu Z, et al. [The Epidemiology of Carbapenemases in Escherichia coli and Klebsiella pneumoniae Isolated in 2019 in Turkey]. Mikrobiyol Bul 2021; 55: 1-16.
  • Çiğdem A, Tuba D, Tuğcan B, Neslihan G, Rıza D. Investigation of carbapenemase and mcr-1 genes in carbapenem-resistant Klebsiella pneumoniae isolates. J Infect Develop Countr 2019; 13: 6.
  • Bilal H, Rehman TU, Khan MA, et al. Molecular Epidemiology of mcr-1, bla (KPC-2,) and bla (NDM-1) Harboring Clinically Isolated Escherichia coli from Pakistan. Infect Drug Resist 2021; 14: 1467-79.
  • Parlak AU, Güdücüoğlu H, Parlak M, Bayram Y, Otlu B. Klebsiella pneumoniaesuşlarında OXA-48 ve alt türevlerinin araştırılması ve fenotipik yansıma. ANKEM Derg 2021; 35: 1-8.
  • Caspar Y, Maillet M, Pavese P, et al. mcr-1 colistin resistance in ESBL-producing Klebsiella pneumoniae, France. Emerg Infect Dis 2017; 23: 874.
  • Robin F, Beyrouthy R, Colot J, et al. MCR-1 in ESBL-producing Escherichia coli responsible for human infections in New Caledonia. J Antimicrob Chemother 2017; 72: 946-7.
  • Gröndahl‐Yli‐Hannuksela K, Lönnqvist E, Kallonen T, et al. The first human report of mobile colistin resistance gene, mcr‐1, in Finland. Apmis 2018; 126: 413-7.
  • Abd El-Baky RM, Masoud SM, Mohamed DS, et al. Prevalence and some possible mechanisms of colistin resistance among multidrug-resistant and extensively drug-resistant Pseudomonas aeruginosa. Infect Drug Resist 2020; 13: 323.
  • Hameed F, Khan MA, Muhammad H, Sarwar T, Bilal H, Rehman TU. Plasmid-mediated mcr-1 gene in Acinetobacter baumannii and Pseudomonas aeruginosa: first report from Pakistan. Revista da Sociedade Brasileira de Medicina Tropical 2019; 52.
There are 27 citations in total.

Details

Primary Language Turkish
Subjects Health Care Administration
Journal Section Research Articles [en] Araştırma Makaleleri [tr]
Authors

Gülnur Tarhan 0000-0002-1019-0798

Funda Şahin 0000-0001-8406-2011

Salih Cesur

Project Number -
Publication Date December 21, 2021
Published in Issue Year 2021

Cite

AMA Tarhan G, Şahin F, Cesur S. Investigation of the presence of OXA 48 and mcr-1 genes in carbapenem-resistant Acinetobacter baumannii, Klebsiella pneumoniae and Pseudomonas aeruginosa strains by in house-PCR method. J Med Palliat Care / JOMPAC / Jompac. December 2021;2(4):118-123. doi:10.47582/jompac.1028959

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