Detection of nosocomial toxigenic Clostridium difficile associated diarrhea in children by conventional PCR

Abstract

Background: Clostridium difficile is a gram positive anaerobic spore forming bacteria. C. difficile–associated disease is a critical clinical issue that is accepted to happen mainly after hospitalization and used of expansive range anti-infection agents. Objective:To define the rate of C. difficile infections isolated from children patients suffering from diarrhea, detection profile toxigenicity of C. difficile strains for toxin A and toxin B by using of PCR, and revise different risk factors of C. difficile infections. Patients and Methods: This cross-sectional study included 50 patients who hospitalized for at least 2 days before the appearance of three or more unformed or liquid stools for 24h, genomic DNA was extracted by using 10% fecal supernatant and a ready kit was used for extraction according to manufacturer instructions. Molecular detection of toxigenic C. difficile done by using the specific primer sequences in polymerase chain reaction. Results: Current study showed diarrhea was the most prominent complain among the study population accounting for 41(82%), of whom 39(78%) presented with watery diarrhea. 38(76%) patients had no fever. The most comorbid disease was inflammatory bowel disease (IBD) with 7 (14%) patients. Forty-six (92%) cases had no history of hospitalization in the last 3 months versus only 8% had such history. PCR revealed that 16 (32%) samples were positive for tcdB gene, while all samples were negative for genes tcdA. Conclusion: The study showed a relationship between previously diagnosed patients with IBD and exacerbations with C. difficile infections (CDIs). Clinically the toxin B alone elicits severe enterotoxic effects which increase rate of (CDIs).