Rola parametrów stanu zapalnego w kale pacjentów z zakażeniem Clostridium difficile: badania wstępne
[The role of inflammatory parameters in feces of patients with Clostridium difficile infections: preliminary study]

STRESZCZENIE

Celem pracy była ocena przydatności oznaczenia kałowej kalprotektyny i laktoferyny w rutynowej, laboratoryjnej diagnostyce zakażeń C. difficile (CDI) i w prognozowaniu przebiegu choroby. Z wyhodowanych z kału 39 pacjentów z CDI 39 szczepów C. difficile, 24 zaklasyfikowano do PCR RT 027, po dwa - do PCR RT 010 i 014, rybotypów 11 szczepów nie ustalono. Dwa izolaty C. difficile PCR RT 027 (8,3%) wykazały oporność na 4 antybiotyki – MDR; w próbkach kału tych pacjentów poziom kalprotektyny przekraczał punkt odcięcia około 7-10 krotnie, a poziom laktoferyny około 24-32 krotnie. Stężenie laktoferyny było  znamiennie wyższe (p<0,005) u pacjentów z dodatnimi wynikami toksyn w kale, jednak nie udało się określić znamienności w przypadku kalprotektyny. Dalsze badania na większej grupie pacjentów z CDI będą przydatne, aby wykazać rolę kalprotektyny i laktoferyny, jako ważnych markerów stanu zapalnego.

ABSTRACT

Introduction: Clostridium difficile infections (CDI), also known as antibiotic-associated

diarrhoea, can result in severe infections in the intestinal tract with systemic complications. A high activity of neutrophils, monocytes and other cells during intestinal infection can result in production of several proteins, including lactoferrin and calprotectin that can be detected in fecal samples. The aim of this study was to evaluate fecal lactoferrin and calprotectin determination in routine diagnostic practice as a predictor for disease progressing.

Methods: Fecal samples of 39 patients suspected for CDI were studied for presence of antigen GDH (C. Quick Quick Check Complete, TechLab, USA) and C. difficile toxins A and/or B (C. diff. Quick Check Complete (TechLab, USA), C. DIFFICILE TOX A/B II ™ (TechLab, USA); and toxin A gene (Illumigene® C. difficile, Meridian Bioscience, USA). In fecal samples lactoferrin (LACTOFERRIN SCAN™; TechLab, USA) and calprotectin levels (Calprest; Eurospital; Italy) were also determined. Fecal samples were cultured for C. difficile, isolated strains were identified, ribotyped and MICs to 11 antibiotics were determined.

Results: Of 39 cultured strains, 24 belonged to PCR RT 027, two strains to each of PCR RT

010 and 014, and 11 strains were not typeable. All isolated C. difficile strains were resistant to ciprofloxacin, and two (multidrug resistant, MDR) strains belonging to PCR RT 027 (8,3%) were resistant to at least 4 antibiotics (clindamycin, erythromycin, moxifloxacin and rifampicin). In fecal samples (n=33) with free C. difficile toxins, lactoferrin level was 0,2 -286,9 μg/mL (median 173,0; IQR=188,1), and calprotectin level 12,8 - 786,8 mg/kg (median 318,5; IQR=220,2). The lactoferrin concentration was significantly higher (p<0,005) in patients with toxin positive tests than in samples without detected toxins, however significant differences were not observed in the calprotectin concentration. Interestingly, fecal samples of two patients infected with MDR strains of PCR RT 027, had calprotectin levels surpassing a cut off between ~7-10 times, similarly as lactoferrin level ~24-32 times.

Conclusions: Further studies on larger group of patients with CDI are required to demonstrate that lactoferrin and calprotectin can be uses as markers of inflammation.

Key words: Clostridium difficile, CDI, calprotectin, lactoferrin