Cultural Isolation and PCR Based Assay for Detection of flaA gene of Campylobacter jejuni from Acute Diarrheic Patients in Tertiary Care Hospital at Dhaka, Bangladesh

Abstract

Aim: Among bacterial causes of acute diarrhea Campylobacter species is frequently responsible. Campylobacter spp. is the leading agents of bacterial gastroenteritis in developed as well as developing countries. This study was conducted to determine the frequency and antimicrobial susceptibility pattern of Campylobacter spp. isolated from stool sample of acute diarrheic patients. Study Design: This was a cross-sectional study. Place and Duration of Study: Two hundred stool samples of children of age between 6 months to 5 years with diarrhea/dysentery were taken from outpatient and inpatient department of Dhaka Medical College Hospital (DMCH) and Dhaka Shishu Hospital from the 1st January, 2011 to 31st December, 2011. The study was carried out at the department of Microbiology, Dhaka Medical College (DMC). Methodology: The samples were inoculated on Campy-Bap media and MacConkey’s agar media and were incubated at 42°C under microaerophilic conditions. The growth after 48 hours was provisionally identified by colonial morphology, oxidase test, catalase test, Gram staining and motility. The organisms were identified to species level by hippurate hydrolysis and resistance to cephalothin. All isolates of Campylobacter jejuni by using conventional bacteriological method were also positive using the Polymerized Chain Reaction (PCR) assay by detecting flaA gene specific for Campylobacter jejuni. Susceptibilities of 27 Campylobacter isolates were determined for seven antimicrobial drugs using the disk diffusion assay. Results: Using conventional bacteriological methods, 27(13.5%) of 200 stool samples were positive for Campylobacter spp. Among isolated Campylobacter spp, Campylobacter jejuni was (88.89%); the remaining isolates were Campylobacter coli (11.11%). Peak age of children with Campylobacter spp. infection was 6-12 months. The male and female ratio was 1.5:1. Resistance to co-trimoxazole was the most common finding, followed by resistance to nalidixic acid, and ciprofloxacin.