Optimization of Eva Green real-time PCR for differentiating C. jejuni/coli directly from feces
OBJECTIVE: To develop and optimize a rapid molecular method for diagnosing campylobacteriosis directly from a clinical fecal sample and at the same time for determining the most common causing agents – C. jejuni/coli.
MATERIALS AND METHODS: 38 clinical fecal samples from hospitalized patients with diarrheal syndrome were tested using a rapid immunochromatographic test. All positive samples were tested for confi rmation by culturing
in a microaerophilic atmosphere. The Eva Green real-time mPCR reaction of a direct fecal sample was conducted using the “IQ5TM Real-Time PCR System” apparatus.
RESULTS: Out of 38 clinical fecal samples which were ICT positive, 18 strains were isolated by culture, namely, 17 of C. jejuni and 1 of C. coli. The Eva Green real-time mPCR reaction also reported 18 positive samples for Campylobacter, out of which 17 were of C. jejuni and only one of C.coli.
CONCLUSION: We developed and optimized the Eva Green real-time mPCR for the detection and species differentiation of C. jejuni/coli directly from a clinical fecal sample. The molecular analysis we described has a 100% sensitivity and specifi city when comparing the results obtained by it to those of the culture method, which is currently the “gold standard” in the diagnosis of campylobacteriosis
KEY WORDS: Campylobacter, diarrhea, mPCR.
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