Abstract

PREVALENCE, ANTIBIOTIC SUSCEPTIBILITY PROFILES AND MOLECULAR METHODS FOR DIAGNOSIS OF CAMPYLOBACTER SPECIES CAUSING CHILDREN GASTROINTESTINAL IN BAHONAR HOSPITALS IN KARAJ

Maryam Shirzadi and Dr. Arman Rostamzad*

ABSTRACT

Introduction: Campylobacter spp. are increasingly being recognized as agents of gastroenteritis worldwide. C. jejuni and C. coli representing the most frequently involved species. Objective: In this survey, the prevalence of campylobacter spp. From diarrheal stools, antibiotic susceptibility profiles, including MIC pattern to different antibiotics and PCR-RFLP technique were determined using the standard microbiological techniques. Methods: Samples were collected from 200 children aged less than 8 years old during May to September of 2013. The routine microbiological methods and polymerase chain reaction (PCR) were performed to diagnosis of Campylobacter strains. The antibiotic susceptibility test was performed using disk diffusion method and MIC of nalidixic acid, imipenem, erythromycin, amoxicillin, gentamicin, tetracycline, doxycycline, vancomycin and kanamycin were determined in microplates and alternatively, the optical density was read in an ELISA reader at 495 nm. The RFLP technique using AluI and TasI restriction enzymes was performed to differentiate C.jejuni, C.coli, C.lari and C.upsaliensis species. Results: Totally 21species (10.5%) detected as Campylobacter spp. Using PCR and bacteriological methods and 11 species (52.38%) were resistant to gentamicin and tetracycline, 10 species (47.61%) were resistant to kanamycin, doxycycline and ciprofloxacin, 12 species(57.14%) were resistant to nalidixic acid and erythromycin, 19 species (90.47%) were resistant to amoxicillin and erythromycin, and 3 species (14.28%) were resistant to imipenem. A wide range of MICs was observed among isolates, and results of PCR-RFLP showed that, 11species (5.5%) showed the Campylobacter jejuni electrophoretic pattern and 7species (3.5%) showed the Campylobacter coli electrophoretic pattern and 3species (1.5%) showed both of the patterns and mixed infection. Conclusion: The PCR-RFLP technique can be used as a rapid, sensitive and specific method for detection and simultaneous differentiation of Campylobacter species in clinical samples.

Keywords: Campylobacter, Children stool, PCR, PCR-RFLP.