DETECTION OF TOXIGENIC CLOSTRIDIUM DIFFICILE IN COLITIS PATIENTS STOOL BY REAL TIME PCR

The 17 th International and Iranian Congress of Microbiology

23 till 25 August 2016، Tehran - Iran

Presentation Type: Speech
Abstract:

Background and Aim: Clostridium difficile is a Gram - positive anaerobic bacillus, with the ability to produce spores and is the etiologic agent of nosocomial pathogens with widespread gastrointestinal diseases as self - limiting diarrhea to fatal colitis caused by antibiotics therapy. Clos tridium. difficile can colonize the large intestine by adhesion of the bacteria to grow and produce toxin type A and toxin type B (TcdA,Tcd B), which leads to the formation of different symptoms of the disease such as pseudomembrane colitis; particularly i n the elderly is one of the most important factors be considered hospital acquired infections. That's why rapid diagnosis of bacteria in patient samples is especially important. Methods: In a cohort study at an academic medical center, samples of colitis pa tients with a history of antibiotic treatment collected. Stool samples cultured on the CCFA media following alcohol shock. Several methods assessed including: culture PCR, stool PCR (cdd - 3, tcdA&tcdB), cell - culture toxin assay, Real - time PCR (tcdA&tcdB) cu ltured isolated and stool specimens. Real - time PCR performed with tcdA&tcdB gene - specific SYBR® Green primers and using Lightcycler® Systems(Roche Diagnostics). Results: A total of 11 stool samples were positive by Real - time PCR. Sensitivity of all methods were 100%, but specificity of culture, culture PCR and culture and stool real - time PCR were 81%, 100% and 100% respectively. Conclusion: Therefore, real - time PCR stool sample has greater value percent with higher specificity. Due to slow growing of C. dif ficile, rapid diagnosis from clinical samples provide effective help for the medical staff.