Optimization of Eva Green real-time mPCR for differentiating C. jejuni/coli directly from feces
| Autor: | E. Dobreva, M. Pavlova, Iskra Tomova, Valeri Velev, Ivan N. Ivanov, P. Petrov, Todor Kantardjiev, G. Asseva, Atanas Mangarov |
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| Rok vydání: | 2017 |
| Předmět: |
DNA
Bacterial Diarrhea Economics and Econometrics Hospitalized patients Immunochromatographic test Campylobacteriosis Campylobacter coli Biology Real-Time Polymerase Chain Reaction medicine.disease_cause Sensitivity and Specificity Microbiology Campylobacter jejuni Feces Campylobacter Infections Materials Chemistry Media Technology medicine Humans Campylobacter Forestry Gold standard (test) medicine.disease Molecular analysis Genes Bacterial |
| Zdroj: | Bratislava Medical Journal. 118:702-704 |
| ISSN: | 1336-0345 |
| DOI: | 10.4149/bll_2017_132 |
| Popis: | 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). |
| Databáze: | OpenAIRE |
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