KLİNİK ÖRNEKLERDEN İZOLE EDİLEN KLEBSİELLA PNEUMONİAE İZOLATLARINDA KARBAPENEM DİRENCİNİ SAPTAMAKTA KULLANILAN FENOTİPİK YÖNTEMLERİN PERFORMANSLARININ KARŞILAŞTIRILMASI
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2024-01
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info:eu-repo/semantics/openAccess
Özet
Karbapenem dirençli Enterobacteriaceae’nın neden olduğu enfeksiyonlar küresel bir tehdit haline gelmiştir. Özellikle çoklu ilaca direnç, çeşitli bakteri türleri arasında endişe verici bir oranda artmakta ve hem hastane kaynaklı hem de toplum kaynaklı enfeksiyonlara sebep olmaktadır. Dünya Sağlık Örgütü “Bakteriyel antimikrobiyal direncin 2019 yılında küresel 1,27 milyon ölümden doğrudan sorumlu olduğu ve 4,95 milyon ölüme katkıda bulunduğu tahmin edilmektedir” şeklinde açıklaması bulunmaktadır. Çoğu zaman tedavinin son seçeneği olan karbapenemlere karşı artan direnç yaygınlaşmaktadır. Karbapenemaz üreten kökenlerin ve aynı zamanda taşıyıcıların da erken ve hızlı tanımlanması, bu yüksek dirençli patojenlerin yayılmasını önlemek için zorunludur. Taşıyıcıların erken tanımlanması ve enfeksiyon kontrol önlemlerinin uygulanması, karbapenemazın neden olduğu hastane kaynaklı salgınları önlemenin tek yoludur ve sınırlı tedavi seçeneği vardır. Bu sebeple karbapenemazların hızlı ve doğru tespiti için moleküler yöntemler ve fenotipik yöntemler kullanılmaktadır. Moleküler yöntemler zahmetli olup, deneyimli personel, ekipman gereksinimi ve maliyet gibi kısıtlamalar söz konusudur. European Committee on Antimicrobial Susceptibility Testing (EUCAST) karbapenem duyarlılığı azaldığında karbapenemaz tespiti için fenotipik testleri önerdiği için biz de iki merkezli çalışmamızda fenotipik yöntemlerin performanslarını karşılaştırıp değerlendirmeyi amaçladık. Çalışmaya Karabük Eğitim ve Araştırma Hastanesi ile Balıkesir Atatürk Şehir Hastanesi Mikrobiyoloji laboatuvarlarına çeşitli kliniklerden gönderilen ve izole edilen 100 K.pneumoniae suşu dahil edilmiştir. Suşların tanımlamaları ve karbapenem duyarlılıkları BD Phoenix 100 (Becton Dickinson, USA) ile yapılmıştır. Çalışmaya dahil edilen suşlarda karbapenemaz varlığını tespit etmek için CIM, sCIM, mCIM ve mCIM-A testleri çalışılmıştır. Çalışmaya dahil edilen kökenlerde karbapenemaz direnç genini tespit etmek için altın standart olarak kabul edilen PCR yöntemi kullanılmıştır. Real time PCR yöntemi ile KPC, NDM, VIM, IMP, OXA-51, OXA-23-58 ve OXA-48 direnç genleri araştırılmıştır. Çalışılan fenotipik testler altın standart yöntem olan PCR ile karşılaştırılarak performansları değerlendirilmiştir. Karabük Eğitim ve Araştırma Hastanesi Mikrobiyoloji laboratuvarına gönderilen ve izole edilen 50 K. pneumoniae suşunda CIM, sCIM, mCIM ve mCIM-A testlerinin özgüllüğü %100; duyarlılıkları ise sırasıyla %90, %94, %92 ve %98 olarak tespit edilmiştir. Balıkesir Atatürk Şehir Hastanesi Mikrobyoloji laboratuvarına gönderilen ve izole edilen 50 K. pneumoniae suşunda ise CIM, sCIM, mCIM ve mCIM-A testlerinin özgüllüğü %100; duyarlılıkları ise sırasıyla %82, %88, %90 ve %90 olarak tespit edilmiştir. Çalışmamızda fenotipik testlerin duyarlılık ve özgüllükleri yüksek oranda saptanmış olup mCIM-A yöntemi karbapenem direncini saptamada en etkin yöntem olarak bulunmuştur
Infections caused by carbapenem-resistant Enterobacteriaceae have become a global threat. Especially multidrug resistance is increasing at a concerning rate among various bacterial species, causing both hospital-acquired and community-acquired infections. The World Health Organization has stated that bacterial antimicrobial resistance was directly responsible for an estimated 1.27 million deaths globally in 2019 and contributed to 4.95 million deaths."" Increasing resistance to carbapenems, often the last treatment resort, is becoming widespread. Early and rapid identification of infected patients and carriers producing carbapenemases is essential to prevent the spread of these highly resistant pathogens. Early identification of carriers and implementation of infection control measures is the only way to prevent hospital-acquired outbreaks caused by carbapenemases, and there are limited treatment options. Therefore, molecular and phenotypic methods are used for the rapid and accurate detection of carbapenemases. ""Molecular methods are subject to limitations such as labor intensiveness, requirement for experienced personnel, equipment, and cost."" Since European Committee on Antimicrobial Susceptibility Testing (EUCAST) recommends phenotypic tests for the detection of carbapenemases, if sensitivity of carbapenem susceptibility tests decreases. Therefore we aimed to compare and evaluate the performance of phenotypic methods in our two-center study. The study included 100 K. pneumoniae strains isolated from various clinics and sent to the Microbiology laboratories of Karabük Training and Research Hospital and Balıkesir Atatürk City Hospital. The strains were identified and their carbapenem susceptibilities were determined using the BD Phoenix 100 (Becton Dickinson, USA)."" CIM, sCIM, mCIM, and mCIM-A tests were performed to detect the presence of carbapenemases in the included strains."" The gold standard test, PCR, considered the gold standard for detecting the resistance gene of each included strain, was performed. The resistance genes KPC, NDM, VIM, IMP, OXA-51, OXA-23, OXA-58, and OXA-48 were investigated using the real-time PCR method. The performance of the studied phenotypic methods was evaluated by comparing them with the gold standard method, PCR. The specificity and sensitivity of CIM, sCIM, mCIM, and mCIM-A tests were 100%, 90%, 94%, 92%, and 98%, respectively, for 50 K.pneumoniae strains isolated from the microbiology laboratory of Karabük Training and Research Hospital. We determined the specificity of CIM, sCIM, mCIM, and mCIM-A tests for the 50 K.pneumoniae strains sent to the Microbiology laboratory of Balıkesir Atatürk City Hospital as 100%, and their sensitivities as 82%, 88%, 90%, and 90% respectively. In our study, the sensitivity and specificity of phenotypic tests were found to be high, and the mCIM-A method was found to be the most effective method in detecting carbapenem resistance."
Infections caused by carbapenem-resistant Enterobacteriaceae have become a global threat. Especially multidrug resistance is increasing at a concerning rate among various bacterial species, causing both hospital-acquired and community-acquired infections. The World Health Organization has stated that bacterial antimicrobial resistance was directly responsible for an estimated 1.27 million deaths globally in 2019 and contributed to 4.95 million deaths."" Increasing resistance to carbapenems, often the last treatment resort, is becoming widespread. Early and rapid identification of infected patients and carriers producing carbapenemases is essential to prevent the spread of these highly resistant pathogens. Early identification of carriers and implementation of infection control measures is the only way to prevent hospital-acquired outbreaks caused by carbapenemases, and there are limited treatment options. Therefore, molecular and phenotypic methods are used for the rapid and accurate detection of carbapenemases. ""Molecular methods are subject to limitations such as labor intensiveness, requirement for experienced personnel, equipment, and cost."" Since European Committee on Antimicrobial Susceptibility Testing (EUCAST) recommends phenotypic tests for the detection of carbapenemases, if sensitivity of carbapenem susceptibility tests decreases. Therefore we aimed to compare and evaluate the performance of phenotypic methods in our two-center study. The study included 100 K. pneumoniae strains isolated from various clinics and sent to the Microbiology laboratories of Karabük Training and Research Hospital and Balıkesir Atatürk City Hospital. The strains were identified and their carbapenem susceptibilities were determined using the BD Phoenix 100 (Becton Dickinson, USA)."" CIM, sCIM, mCIM, and mCIM-A tests were performed to detect the presence of carbapenemases in the included strains."" The gold standard test, PCR, considered the gold standard for detecting the resistance gene of each included strain, was performed. The resistance genes KPC, NDM, VIM, IMP, OXA-51, OXA-23, OXA-58, and OXA-48 were investigated using the real-time PCR method. The performance of the studied phenotypic methods was evaluated by comparing them with the gold standard method, PCR. The specificity and sensitivity of CIM, sCIM, mCIM, and mCIM-A tests were 100%, 90%, 94%, 92%, and 98%, respectively, for 50 K.pneumoniae strains isolated from the microbiology laboratory of Karabük Training and Research Hospital. We determined the specificity of CIM, sCIM, mCIM, and mCIM-A tests for the 50 K.pneumoniae strains sent to the Microbiology laboratory of Balıkesir Atatürk City Hospital as 100%, and their sensitivities as 82%, 88%, 90%, and 90% respectively. In our study, the sensitivity and specificity of phenotypic tests were found to be high, and the mCIM-A method was found to be the most effective method in detecting carbapenem resistance."
Açıklama
Anahtar Kelimeler
Karbapenem direnci, CIM (Karbapenem İnaktivasyon Metodu), mCIM (Modifiye Karbapenem İnaktivasyon Metodu), sCIM (Basitleştirilmiş Karbapenem İnaktivasyon Metodu), mCIM-A (Amonyum bikarbonat ilaveli Modifiye Karbapenem İnaktivasyon Metodu), Carbapenem Resistance, CIM (Carbapenem Inactivation Method), mCIM (Modified Carbapenem Inactivation Method), sCIM (Simplified Carbapenem Inactivation Method), mCIM-A (Modified Carbapenem Inactivation method with ammonium bicarbonate)