Molecular Detection of Clostridium Difficile in Patients with Diarrhea via LAMP Technique

Background and Clostridium difficile is an obligate anaerobic, gram positive bacillus. The purpose of this study was to develop and optimize Loop-mediated Isothermal Amplification (LAMP) technique for assessing the prevalence of Clostridium difficile infection in the samples of watery diarrhea. This cross-sectional qualitative study was performed in 48 samples of watery diarrhea (fecal samples were obtained from 24 patients with nosocomial diarrhea and 24 patients with antibiotic-associated diarrhea). All samples were cultured in the specialized CDMN Agar medium. Six specific primers were applied for the LAMP Assay and the LAMP reaction was conducted under optimal conditions. Furthermore, sensitivity and specificity of LAMP were determined to detect Clostridium difficile. In this study, the optimized LAMP technique was determined to be 10 bacterium. In addition, LAMP technique showed a high specificity for detecting the bacteria. Among the patients with nosocomial diarrhea and antibiotic-associated diarrhea, 18.2% and11.5% were reported to be positive, respectively. LAMP was found as an applicable, sensitive, and quick technique for detection of Clostridium difficile.