Investigation of Clostridioides (Clostridium) difficile in patients with hospital-acquired diarrhea

Öz

Aim: Clostridioides difficile infection is a healthcare-associated infection and is an important source of mortality and morbidity, especially for hospitalized patients. In our study, we aimed to determine the incidence of C. difficile in hospital-acquired diarrhea by isolating C. difficile and investigating the presence of toxin in stool samples of hospitalized patients. Method: The study included 85 patients over two years of age who had loose or watery stools more than three times a day for at least two days and hospitalized in various wards at Mersin University Hospital between October 2020 and July 2021. Stool samples collected in sealed containers were analyzed for the presence of C. difficile toxin A/B using the 'Epilisa Fecal C. difficile Toxin A/B ELISA Kit' (Epitope Diagnostics, Inc. ABD) within 24-48 hours. In addition, stool samples were cultured on C. difficile agar containing cycloserine, cefoxitin, amphotericin-B (Biomerieux, Fransa), ChromID C. difficile agar (Biomerieux, Fransa) and 5% sheep blood Columbia agar (Biomerieux, Fransa) media. The media were incubated in an anaerobic environment at 37°C for 48-72 hours and then evaluated. The colonies identified as C. difficile were inoculated on Schaedler Anaerobic Broth (Oxoid, İngiltere) and after 48-72 hours of anaerobic incubation, the presence of toxin was investigated by ELISA method. Results: ELISA toxin A/B positivity was found to be 4.7% (4/85) in our study. Six (7.1%) C. difficile isolates were isolated in anaerobic culture and the presence of toxin was detected in three of them (p=1.000). The frequency of CDE in our patient population was determined to be 3.5% (n=3). When toxigenic culture was taken as the reference test, the sensitivity, specificity, positive predictive value and negative predictive value of ELISA Toxin A/B test were determined as 100%, 98.8%, 75% and 100%, respectively. Conclusion: Rational antibiotic use and appropriate infection control programs should be considered in the prevention of CDE, which is associated with antibiotic use and hospitalization

Anahtar Kelimeler

• Hospital-acquired
• diarrhea
• Clostridium difficile
• toxin A/B
• anaerobic culture

Hastane kaynaklı ishallerde Clostridioides (Clostridium) difficile araştırılması

Öz

Amaç: Clostridioides difficile enfeksiyonu, sağlık hizmeti ile ilişkili bir enfeksiyon olup özellikle hastanede yatan hastalar için önemli mortalite ve morbidite kaynağıdır. Çalışmamızda; hastane kaynaklı ishallerde C. difficile insidansını belirlemek için yatan hastaların gaita örneklerinde C. difficile izolasyonu ve toksin varlığının araştırılması amaçlanmıştır. Yöntem: Çalışmaya, Ekim 2020-Temmuz 2021 tarihleri arasında Mersin Üniversitesi Hastanesi’nde çeşitli servislere yatırılan, en az iki gün boyunca günde üçten fazla gevşek veya sulu dışkılama yapan, iki yaş üzerinde 85 hasta dahil edilmiştir. Sızdırmaz kaplarda toplanan gaita örneklerinde, 24-48 saat içinde ‘Epilisa Fecal C. difficile Toksin A/B ELISA kiti (Epitope Diagnostics, Inc. ABD) kullanılarak C. difficile toksin A/B varlığı araştırılmıştır. İlave olarak gaita örnekleri; sikloserin, sefoksitin, amfoterisin-B içeren C. difficile agar (Biomerieux, Fransa), ChromID C. difficile agar (Biomerieux, Fransa) ve %5 koyun kanlı Columbia agar (Biomerieux, Fransa) besiyerlerine ekilmiştir. Besiyerleri 37°C’lik etüvde anaerop ortamda 48-72 saat inkübe edildikten sonra değerlendirilmiştir. C. difficile olarak tanımlanan kolonilerden Schaedler Anaerop buyyona (Oxoid, İngiltere) ekim yapılmış ve 48-72 saatlik anaerop inkübasyonun sonunda ELISA yöntemiyle toksin varlığı araştırılmıştır. Bulgular: Çalışmamızda ELISA toksin A/B pozitifliği %4.7 (4/85) olarak bulunmuştur. Anaerop kültürde üreyen altı (%7.1) C. difficile izolatının üçünde toksin varlığı saptanmıştır (p=1.000). Hasta popülasyonumuzda CDE sıklığının %3.5 (n=3) olduğu belirlenmiştir. Toksijenik kültür referans test olarak alındığında, ELISA Toksin A/B testinin duyarlılık, özgüllük, pozitif prediktif değer ve negatif prediktif değeri sırasıyla; %100, %98.8, %75 ve %100 olarak belirlenmiştir. Sonuç: Antibiyotik kullanımı ve hastane yatışı ile ilişkili olan CDE’nun önlenmesinde akılcı antibiyotik kullanımı ve uygun enfeksiyon kontrol programlarının göz önünde bulundurulması gereklidir.

Anahtar Kelimeler

• Hastane kaynaklı
• ishal
• Clostridium difficile
• toksin A/B
• anaerobik kültür

Kaynakça

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