Çeşitli Klinik Örneklerden İzole Edilen Pseudomonas aeruginosa'nın Karbapenemaz Üretimi ve Tiplendirilmesinde Fenotipik ve Genotipik Yöntemlerin Değerlendirilmesi

Year 2024, Volume: 4 Issue: 2, 42 - 49, 30.05.2024

Hatice Karali , Mehmet Mücahit Güncü , Mehmet Burak Aksu

Abstract

Amaç: Son on yılda karbapenem dirençli Pseudomonas aeruginosa izolatlarının artışı enfeksiyon tedavisinde önemli bir sağlık sorunu haline gelmiştir. Rutin laboratuvarda karbapenem direncinin saptanması için gereken süre yaklaşık 48 saattir. Bu durum, özellikle kritik hastalarda etkili tedavinin başlatılmasında gecikmeye neden olur. Bu nedenle çalışmalar antimikrobiyal direncin daha erken tespit edilmesi için yeni yöntemlere odaklanmaktadır. Bu çalışmada, karbapenem dirençli P. aeruginosa izolatlarında karbapenemaz üretimine bağlı direncin fenotipik ve genotipik yöntemlerle araştırılması amaçlanmıştır.
Yöntemler: Marmara Üniversitesi Pendik Eğitim ve Araştırma Hastanesi Mikrobiyoloji Laboratuvarı’nda Ocak 2018 ile Şubat 2023 tarihleri arasında çeşitli klinik örneklerden izole edilen 120 P. aeruginosa izolatı rutin antibiyogram sonuçlarına bakılarak çalışmamıza dahil edilmiştir. Karbapenemaz üretimi fenotipik test olarak enzim-substrat etkileşimine dayalı reaksiyon temelli kolormatik yöntem ve genotipik test olarak polimeraz zincir reaksiyonu ile tespit edilmiştir.
Bulgular: Bu çalışmada karbapenem dirençli 100 izolatın 46'sında (%46) genotipik yöntemle karbapenemaz kodlayan genler tespit edilmiştir. Fenotipik test ile karbapenemaz enzimi taşıyan 46 izolatın 31’inde (%67) 1 saat içinde pozitif sonuçlar kaydedilmiştir. Geriye kalan 13 izolat yanlış negatif olarak; moleküler yöntem ile direnç genlerini taşımadığı belirlenen 2 izolat ise yanlış pozitif olarak değerlendirilmiştir. Fenotipik testin duyarlılık ve özgüllüğü sırasıyla; %67.4 ve %97.3 (p<0.0001) olarak bulunmuştur.
Sonuç: Sonuç olarak, karbapenem dirençli P. aeruginosa izolatlarının hızlı ve doğru tanımlanması, zamanında uygun tedavinin verilmesi ve enfeksiyon kontrol önlemlerinin başarılı bir şekilde uygulanması açısından oldukça önemlidir. Çalışmamızda kullandığımız enzim-substrat etkileşimine dayalı reaksiyon temelli kolormatik testin performansının değerlendirilebilmesi için daha fazla örnek içeren in vitro çalışmalara ihtiyaç vardır.

Keywords

Pseudomonas aeruginosa, antibiyotik direnci, karbapenem, karbapenemaz, fenotipik test

Ethical Statement

Bu çalışma, Marmara Üniversitesi Tıp Fakültesi Klinik Araştırmalar Etik Kurulu tarafından (03.02.2023 tarih ve 265 kayıt numarası) onaylanmıştır.

Supporting Institution

Marmara Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi (BAPKO)

Project Number

TYL-2023-11039

Evaluation of Phenotypİc and Genotypic Methods for Carbapenemase Production and Typing In Pseudomonas aeruginosa Isolates from Various Clinical Samples

Abstract

Objective: The increase in carbapenem-resistant Pseudomonas aeruginosa isolates has become an important health problem in infection treatment in the last decade. The time required to detect carbapenem resistance in the routine laboratory setting is about 48 hours. So, it causes a delay in the initiation of effective treatment, especially in critically ill patients. For this reason, studies focus on new methods to detect antimicrobial resistance earlier. It aimed to investigate carbapenemase-dependent resistance by phenotypic and genotypic methods in carbapenem-resistant clinical P. aeruginosa isolates in this study.
Methods: A hundred twenty P. aeruginosa isolates obtained from clinical samples between January 2018 to February 2023 in the Microbiology Laboratory of Marmara University Pendik Training and Research Hospital, were included in our study based on routine antibiogram results. Carbapenemase production was detected by enzyme-substrate reaction-based colorimetric method as a phenotypic test and polymerase chain reaction as a genotypic test.
Results: In this study, carbapenemase-coding genes were detected in 46 (46%) of 100 carbapenem-resistant isolates by genotypic method. With the phenotypic test, positive results were recorded within 1 hour in 31 of 46 isolates (67%) carrying the carbapenemase enzyme. The remaining 13 isolates were false negatives; 2 isolates determined not to carry the resistance genes by molecular method were evaluated as false positives. The sensitivity and specificity of the phenotypic test were 67.4% and 97.3%, respectively (p<0.0001).
Conclusion: In conclusion, rapid and accurate identification of carbapenem-resistant P. aeruginosa isolates is very important for the timely administration of appropriate treatment and successful implementation of infection control measures. In vitro studies with a larger number of samples are needed to evaluate the performance of the enzyme-substrate reaction-based colorimetric test that we used in our study.

Keywords

Pseudomonas aeruginosa, antibiotic resistance, carbapenem, carbapenemase, phenotypic test

Project Number

TYL-2023-11039

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