Ocena stężenia kalprotektyny kałowej w próbkach pacjentów z zakażeniem Clostridium difficile
[Assessment of fecal calprotectin concentration in samples from patients with Clostridium difficile infection]

STRESZCZENIE

Celem pracy była ocena stężenia kalprotektyny w anonimizowanych, resztkowych pięćdziesięciu ośmiu próbkach kału, w których wykryto markery Clostridium difficile oraz ustalenie związku między stężeniem kalprotektyny a typem genetycznym (PCR-rybotypem (RT) C. difficile. Z próbek wyhodowano 47 toksynotwórczych szczepów C. difficile, które zakwalifikowano, stosując metodę PCR-rybotypowanie, do następujących pięciu PCR-rybotypów (RT): RT027 (n=39), RT020 (n=4), RT176 (n=4), RT001 (n=1) i RT017 (n=1). Stężenie kalprotektyny w resztkowych próbkach kału było w zakresie od 199,7-1137,1 μg/g kału (wartości >150), co wskazywało na toczący się proces zapalny. Stwierdzono wysokie stężenie kalprotektyny w próbkach kału, natomiast nie wykazano statystycznie istotnych związków między wysokim stężeniem kalprotektyny a PCR-rybotypem C. difficile (p >0,05).

ABSTRACT

Introduction: Clostridium difficile infection (CDI) can cause of nosocomial diarrhoea and intestinal inflammation. Fecal calprotectin (fc) is a sensitive marker of intestinal inflammation, found to be associated with enteric bacterial infections as C. difficile infection (CDI).
Materials and Methods: We evaluated faecal calprotectin (fc) levels using a Ridascreen® Calprotectin test (R-Biopharm AG) in faecal samples of CDI positive patients diagnosed by C. Diff Quik Chek Complete (TechLab; Blacksburg, VA, USA and Alere; Waltham, MA, USA) test. Statistical analysis was performed for correlation of fc levels with infection with different C. difficile PCR-ribotypes (RTs). The study involved two groups of faecal samples: 58 samples with glutamate dehydrogenase (GDH) plus TOX A/B positive and second group were 17 samples from healthy adults without diarrhoea and GDH/TOX A/B negative. All samples (n=75) were plated on CLO selective agar (bioMérieux, Marcy l’Etoile, France) to grow a C. difficile strain. RTs was defined using specific primers.
Results: From 75 stool samples, 47 strains of C. difficile were cultured only from diarrheal patients. In 58 diarrhoeic stool samples, the range of calprotectin concentrations were 199.7-1137.15 μg/g (>150μg/g, elevated concentration), which indicates on inflammatory process. C. difficile strains belonged to the following RTs: RT 027 (n=39), RT 020 (n=4), RT 176 (n=2), RT 001 (n=1) and RT 017 (n=1). In the 17 samples from the healthy adults, the range of calprotectin concentrations was 12.1-25.3 μg/g stool (FC normal concentration C. difficile strain was not cultured.
Conclusions: The study found a differentiated level of calprotectin in the gastrointestinal tract in patients with CDI. There were no statistically significant differences in the concentration of calprotectin between patients infected with RT027 and different PCR ribotypes than RT027 (p >0,05). However, it should be emphasized that these tests carried out on a small number of samples. Examination of relationship between level of calprotectin and genetic type of C. difficile causing CDI requires further investigation.

Key words: calprotectin, Clostridium difficile infection (CDI), PCR- ribotype