Bir üniversite hastanesinde karbapenem dirençli klebsiella pneumoniae suşlarında karbapenem direnç genlerinin moleküler analizi ve suşların klonal ilişkisi
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Dosyalar
Tarih
2017
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Cilt Başlığı
Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Klebsiella pneumoniae önemli insan patojenlerinden biridir. Özellikle hastane kaynaklı enfeksiyonlarda saptanan suşlarında gözlenen karbapenem direnci tedavi alternatiflerini önemli düzeyde kısıtlamaktadır. Klebsiella türlerinde karbapenem direncinin gelişiminin önlenmesi ve dirençli suşların yayılımının azaltılması için uygulanacak önlemlerin geliştirilmesinde bu dirence neden olan moleküler dinamiklerin bilinmesi önem taşımaktadır. Bu çalışmada, klinik örneklerde saptanan karbapenem dirençli K. pneumoniae suşlarında daha önce tanımlanmış olan karbapenemaz genlerinin araştırılması ve izolatlar arasındaki klonal ilişkinin saptanması amaçlanmıştır. Gereç ve Yöntem: Kesitsel bir araştırma olarak planlanan bu çalışmaya hastanemiz tıbbi mikrobiyoloji laboratuvarına gönderilen çeşitli klinik örneklerden Ocak 2012-Haziran 2014 tarihleri arasında soyutlanan karbapenem dirençli K. pneumoniae suşları dahil edilmiştir. Prospektif olarak toplanan izolatların karbapenem direnci ertapenem disk yöntemi ve Modifiye Hodge testi çalışılmış; izolatların imipenem MİK düzeyleri Etest yöntemi ile saptanmıştır. Suşlarda OXA-48, NDM, VIM, IMP, KPC karbapenemaz genleri in-house PZR ile ve izolatlar arasındaki klonal ilişki Pulsed Field Gel Electrophoresis (PFGE) yöntemi ile araştırılmıştır. Bulgular: Çalışma süresince 70 karbapenem dirençli K. pneumoniae suşu tanımlanmıştır. Yapılan moleküler analizde toplam 61 (%87.1) izolatta en az bir karbapenemaz geni saptanmış geri kalan 9 (%12.9) suşta ise çalışılan genlerden herhangi biri bulunamamıştır. Çalışma sonucunda suşların 51'inde (%72.8) blaOXA-48, 11'inde (%15.7) blaNDM, 3'ünde (%4.2) blaVIM bulunmuş ve blaOXA-48 üreten suşlardan 4'ünde (%7.8) blaNDM gen birlikteliği olduğu gözlenmiştir. Çalışmaya dahil edilen suşların hiçbirinde blaIMP ve blaKPC saptanmamıştır. Yapılan Etest MİK analizinde, çalışmaya alınan 70 izolatın 23'ünün (%32,8) imipenem MİK düzeyi duyarlı sınırlar içinde (<4 μg/ml) bulunurken saptanan dağılım 0.125-32 μg/ml arasında olmuştur. Toplam 70 K. pneumoniae suşundan 67'si 59 farklı PFGE profili göstermiş ve geri kalan 3 suş ise anlamlı bir bant profili oluşturmamıştır. Klonal yönden ilişkili suşlar, 4 farklı küme içerisinde yer almış ve suşların kümeleşme oranı 69 %18 olarak saptanmıştır. En büyük küme beş üyeli genotip 58 olup bunu sırayla 3 üyeli genotip 34 ve 2 üyeli genotip 29 takip etmiştir. Tartışma ve Sonuç: Bu çalışmada elde ettiğimiz sonuçlara göre, OXA-48 ve NDM türü karbapenemazlar hastanemizdeki Klebsiella suşlarında görülen karbapenem direncinin çok büyük bir bölümünden sorumludur. Bu bulgular Ülkemizden daha önce bildirilen veriler ile oldukça benzerlik göstermektedir. Bu çalışma tek bir merkezde yapılmış olmasına rağmen suşların oldukça heterojen bir genotipik dağılım profili göstermesi, bu direnç genlerinin farklı kökenler arasında yayılım gösterdiğinin en önemli bulgusu olmuştur.
Aim: Klebsiella pneumoniae is among the leading human pathogens. The carbapenem resistance, particularly seen in the strains from the nosocomial infections, significantly limits the treatment options. Knowing the molecular dynamics of this resistance is important for development of the control measurements intended to prevent the emergence of carbapenem resistance in Klebsiella strains and to reduce the spreading of such resistant strains. In this study, previously determined carbapenemase genes and the clonal relationships of the carbapenem resistant K. pneumonia strains isolated from clinical samples were aimed to be investigated. Material and Methods: In this cross sectional study, the carbapenem resistant K. pneumoniae strains which were isolated from the clinical samples between January 2012 and June 2014 in the medical microbiology laboratory of our hospital were included. Strains were prospectively collected, the carbapenem resistance was determined with ertapenem disc and Modified Hodge test, and imipenem MICs of these strains were studied with Etest method. Carriage of the carbapenemase genes including OXA-48, NDM, VIM, IMP, and KPC were determined with in-house PCR, and clonal relationships of the strains was studied with Pulsed Field Gel Electrophoresis (PFGE). Results: AAim: Klebsiella pneumoniae is among the leading human pathogens. The carbapenem resistance, particularly seen in the strains from the nosocomial infections, significantly limits the treatment options. Knowing the molecular dynamics of this resistance is important for development of the control measurements intended to prevent the emergence of carbapenem resistance in Klebsiella strains and to reduce the spreading of such resistant strains. In this study, previously determined carbapenemase genes and the clonal relationships of the carbapenem resistant K. pneumonia strains isolated from clinical samples were aimed to be investigated. Material and Methods: In this cross sectional study, the carbapenem resistant K. pneumoniae strains which were isolated from the clinical samples between January 2012 and June 2014 in the medical microbiology laboratory of our hospital were included. Strains were prospectively collected, the carbapenem resistance was determined with ertapenem disc and Modified Hodge test, and imipenem MICs of these strains were studied with Etest method. Carriage of the carbapenemase genes including OXA-48, NDM, VIM, IMP, and KPC were determined with in-house PCR, and clonal relationships of the strains was studied with Pulsed Field Gel Electrophoresis (PFGE). Results: A total 70 carbapenem resistant K. pneumoniae were identified throughout the study. With the molecular analysis, at least one carbapenemase gene was determined in 61 (87.1%) strains, and there was no studied genes in any of the remaining 9 (12.9%) strains. After the analyses, blaOXA-48 was found in 51 (72.8%) strains, blaNDM in 11 (15.7%) strains, and blaVIM in 3 (4.2%) strains; and it was observed that 4 (7.8%) strains harboring blaOXA-48 were also carrying blaNDM. blaIMP and blaKPC were not determined any of the studied strains. With the Etest MIC analyses, imipenem MICs of 23 (32.8%) of the 70 studied strains were found under the susceptibility breakpoint (<4 μg/ml), and total MIC distribution of the included strains was between 0.125-32 μg/ml. A total 67 of 70 studied K. pneumoniae strains showed 59 different PFGE profile and remaining 3 strains didn't produce a meaningful band pattern. Clonally related strains were collected in 4 different cluster, and the clustering rate of all strains was found as 18%. The largest cluster was genotype 58 consisting 5 strains, followed by genotype 34 consisting 3 strains and genotype 29 including 2 strains.Discussion and Conclusion: According to results of this study, OXA-48 and NDM type carbapenemases were responsible for the majority of carbapenem resistance in the Klebsiella strains in our hospital. This data is very similar to the data previously reported from our country. Though this study was performed in a single center, the strains showing a highly heterogenic genetic distribution was a significant evidence of proving that these resistance genes spreading between different genetic clones.
Aim: Klebsiella pneumoniae is among the leading human pathogens. The carbapenem resistance, particularly seen in the strains from the nosocomial infections, significantly limits the treatment options. Knowing the molecular dynamics of this resistance is important for development of the control measurements intended to prevent the emergence of carbapenem resistance in Klebsiella strains and to reduce the spreading of such resistant strains. In this study, previously determined carbapenemase genes and the clonal relationships of the carbapenem resistant K. pneumonia strains isolated from clinical samples were aimed to be investigated. Material and Methods: In this cross sectional study, the carbapenem resistant K. pneumoniae strains which were isolated from the clinical samples between January 2012 and June 2014 in the medical microbiology laboratory of our hospital were included. Strains were prospectively collected, the carbapenem resistance was determined with ertapenem disc and Modified Hodge test, and imipenem MICs of these strains were studied with Etest method. Carriage of the carbapenemase genes including OXA-48, NDM, VIM, IMP, and KPC were determined with in-house PCR, and clonal relationships of the strains was studied with Pulsed Field Gel Electrophoresis (PFGE). Results: AAim: Klebsiella pneumoniae is among the leading human pathogens. The carbapenem resistance, particularly seen in the strains from the nosocomial infections, significantly limits the treatment options. Knowing the molecular dynamics of this resistance is important for development of the control measurements intended to prevent the emergence of carbapenem resistance in Klebsiella strains and to reduce the spreading of such resistant strains. In this study, previously determined carbapenemase genes and the clonal relationships of the carbapenem resistant K. pneumonia strains isolated from clinical samples were aimed to be investigated. Material and Methods: In this cross sectional study, the carbapenem resistant K. pneumoniae strains which were isolated from the clinical samples between January 2012 and June 2014 in the medical microbiology laboratory of our hospital were included. Strains were prospectively collected, the carbapenem resistance was determined with ertapenem disc and Modified Hodge test, and imipenem MICs of these strains were studied with Etest method. Carriage of the carbapenemase genes including OXA-48, NDM, VIM, IMP, and KPC were determined with in-house PCR, and clonal relationships of the strains was studied with Pulsed Field Gel Electrophoresis (PFGE). Results: A total 70 carbapenem resistant K. pneumoniae were identified throughout the study. With the molecular analysis, at least one carbapenemase gene was determined in 61 (87.1%) strains, and there was no studied genes in any of the remaining 9 (12.9%) strains. After the analyses, blaOXA-48 was found in 51 (72.8%) strains, blaNDM in 11 (15.7%) strains, and blaVIM in 3 (4.2%) strains; and it was observed that 4 (7.8%) strains harboring blaOXA-48 were also carrying blaNDM. blaIMP and blaKPC were not determined any of the studied strains. With the Etest MIC analyses, imipenem MICs of 23 (32.8%) of the 70 studied strains were found under the susceptibility breakpoint (<4 μg/ml), and total MIC distribution of the included strains was between 0.125-32 μg/ml. A total 67 of 70 studied K. pneumoniae strains showed 59 different PFGE profile and remaining 3 strains didn't produce a meaningful band pattern. Clonally related strains were collected in 4 different cluster, and the clustering rate of all strains was found as 18%. The largest cluster was genotype 58 consisting 5 strains, followed by genotype 34 consisting 3 strains and genotype 29 including 2 strains.Discussion and Conclusion: According to results of this study, OXA-48 and NDM type carbapenemases were responsible for the majority of carbapenem resistance in the Klebsiella strains in our hospital. This data is very similar to the data previously reported from our country. Though this study was performed in a single center, the strains showing a highly heterogenic genetic distribution was a significant evidence of proving that these resistance genes spreading between different genetic clones.
Açıklama
Anahtar Kelimeler
Mikrobiyoloji, Microbiology
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Özdemir, H. (2017). Bir üniversite hastanesinde karbapenem dirençli klebsiella pneumoniae suşlarında karbapenem direnç genlerinin moleküler analizi ve suşların klonal ilişkisi. Yayımlanmış Uzmanlık Tezi, İnönü Üniversitesi, Malatya.
