Home Bratislava Medical Journal 2017 Bratislava Medical Journal Vol.118, No.11, p.702-704, 2017

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Published Monthly, in English
Founded: 1919
ISSN 0006-9248
(E)ISSN 1336-0345
 

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Bratislava Medical Journal Vol.118, No.11, p.702-704, 2017

Title: Optimization of Eva Green real-time mPCR for differentiating C. jejuni/coli directly from feces
Author: M. Pavlova, V. Velev, E. Dobreva, G. Asseva, I. Ivanov, P. Petrov, A. Mangarov, I. Tomova, T. Kantardjiev

Abstract: 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 confirmation 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 specificity when comparing the results obtained by it to those of the culture method, which is currently the “gold standard” in the diagnosis of campylobacteriosis (Tab. 2, Fig. 1, Ref. 6).

Keywords: Campylobacter, diarrhea, mPCR
Published online: 05-Dec-2017
Year: 2017, Volume: 118, Issue: 11 Page From: 702, Page To: 704
doi:10.4149/BLL_2017_132


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