Prevalence, Molecular Characterization, Pathogenicity and Antimicrobial Susceptibility Pattern of Clostridioides Difficile From Clinical Samples in South Eastern Nigeria.

The prevalence, molecular characterization, pathogenicity and antibioticsensitivity pattern of Clostridioides difficile from clinical samples in South-Eastern Nigeria wereevaluated in this study. Stool samples (450) were collected from patientsinsome selected public and private-owned clinics in the five South-Eastern states. Standard bacteriological methods were employed for enumeration of total anaerobes. Culture of Clostridioides difficile isolates were performed on Clostridium difficile differential agar (supplemented with C. difficile selective supplement). Biochemical tests like catalase, oxidase and reverse CAMP test were conducted before extraction of genomic bacterial DNA for suspected C. difficile isolates. Purification and amplification of bacterial DNA was carried out on 2% agarose gel. Amplified bacteria DNA was sequenced and blasted on the National Centre for Biotechnology Information (NCBI) website. Antibiotics susceptibility was carried out for C. difficile isolates using the Kirby-Bauer disc diffusion technique. PCR technique was employed for the detection of virulence ( tcdA, tcdB, cdtA and cdtB ) and resistance genes ( tetS, tetA and ermB ) in C. difficile isolates. Anaerobe counts for stool samples obtained showed that samples had a mean count of 5.63±0.09 log ₁₀ CFU/g (Anambra state) while counts of 5.61±0.11 log ₁₀ CFU/g and 5.65±0.07 log ₁₀ CFU/g were obtained from Ebonyi and Enugu States. The antibacterial sensitivity revealed that the isolates were sensitive to gentamicin, meropenem and amoxicillin-clavulanic acid. The isolates were resistant to tetracyclines and erythromycin, and the multiple antibiotic resistance index of theisolates showed that the multi-drug resistant isolates had a MAR index of 0.44. All (14 C. difficile isolates) (100%) of C. difficile isolates were found to possess tcdB genes, and 69.56% expressed tcdA genes. One (4.34%) of the C. difficile isolatespossessed cdtA and cdtB genes for binary toxin production.