Introduction: Campylobacter is the most commonly reported enteric bacterial pathogen in humans. Detection of Campylobacter species in microbiology laboratory is based on culture of stool samples on selective media followed by phenotypic identification, which are time consuming and esdo not allow an early treatment. This has emphasized the use of more rapid and accurate detection methodologies than that of slow culture-based techniques. Objective: to compare different methods (culture, multiplex real time PCR and immuno - chromatography) for rapid detection of thermophilic Campylobacter species in stool samples from pediatric patients with gastroenteritis and to determine the antibiotic susceptibility of the isolates. Materials & methods: 100 infant & child ≤ 5 year of age with Campylobacter diarrhea like symptoms were included in the study. Stool samples were collected, transported and divided into 2 aliquots; the first for bacteriologic examination (isolation of Campylobacter on (m-CCDA) supplemented with cefoperazone & amphotericin B selective supplement) and immune-chromatography (RIDA QUICK Campylobacter test) and the second part for DNA extraction and multiplex real time PCR examination. Susceptibility of Campylobacters isolates to different antibiotics was carried out using the disk diffusion method. Results: From the 100 stool samples examined, 22 (22%) were positive for Campylobacter by conventional culture method. Taking culture as the reference method, the sensitivity of multiplex real time PCR was 95.5%, specificity was 100%, PPV and NPV values were 100% and 98.7% respectively. One sample tested negative by multiplex real time PCR but positive by culture (one false negative). C. jejuni was the most commonly isolated Campylobacter species (72.7%) followed by C. coli (27.3%). However, RIDA QUICK test could detect 24/100 (24%) as positive samples, among those 24 positives; 22 were considered true positives and 2 were false results. The PPV for RIDA QUICK test was 91.7% and the NPV was 100%, while the sensitivity was100% and the specificity was 97.4%. Conclusion: Multiplex real time PCR assay is a rapid, simple and practical tool for identification of Campylobacter species commonly associated with pediatric gastroenteritis. Besides being sensitive and specific, it can save time and money because several loci can be amplified simultaneously and so offers an effective alternative to conventional culture method. If not available, we recommend the use of the screening ICT in conjunction with culture, which is rapid, easy and applicable test for the diagnosis of gastroenteritis caused by C. jejuni and C. coli, reducing significantly the time to result to only15 minutes. |
