Molecular Identification of Campylobacter Species

The isolates were confirmed as C. jejuni or C. coli by using a multiplex PCR (mPCR) assay [17 (link)]. flaA-restriction fragment length polymorphism (RFLP) analysis was done as previously described [6 (link)]. The flaA amplicon was digested for 18 h at 37 °C with DdeI (Roche Diagnostics GmbH). The DNA segments were separated using 2.5 % agarose gels (Starlab GmbH, Hamburg, Germany) in tris-borate-EDTA buffer at 200 V for 1 h, stained with ethidium bromide and visualized under UV light. Documentation was done using a Bio Imaging System (Syngene, Cambridge, UK).

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Ahmed M.F., El-Adawy H., Hotzel H., Tomaso H., Neubauer H., Kemper N., Hartung J, & Hafez H.M. (2016). Prevalence, genotyping and risk factors of thermophilic Campylobacter spreading in organic turkey farms in Germany. Gut Pathogens, 8, 28.

Publication 2016

Bio Imaging System

Agarose Assay Diagnostics Ethidium bromide Gels Multiplex pcr Restriction fragment length polymorphism Tris borate edta buffer Uv light

Corresponding Organization : University of Veterinary Medicine Hannover, Foundation

Other organizations : Friedrich-Loeffler-Institut, Freie Universität Berlin

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Variable analysis

independent variables

Multiplex PCR (mPCR) assay used to confirm the isolates as Campylobacter jejuni or Campylobacter coli

dependent variables

Identification of Campylobacter species (Campylobacter jejuni or Campylobacter coli)
FlaA-RFLP analysis

control variables

Time of restriction enzyme (DdeI) digestion (18 h at 37 °C)
Agarose gel electrophoresis conditions (2.5% agarose gels, Tris-borate-EDTA buffer, 200 V for 1 h)
Staining with ethidium bromide and visualization under UV light
Documentation using a Bio Imaging System

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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