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Title: Screening and rapid identification of Campylobacter spp. DNA by FlaA PCR based method on chicken and human fecal samples in Egypt

Author
item RAMADAN, HAZEM - Mansoura University
item Jackson, Charlene
item Hinton Jr, Arthur

Submitted to: International Journal of Poultry Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/10/2015
Publication Date: 6/30/2015
Citation: Ramadan, H., Jackson, C.R., Hinton Jr, A. 2015. Screening and rapid identification of Campylobacter spp. DNA by FlaA PCR based method on chicken and human fecal samples in Egypt. International Journal of Poultry Science. 14(5):252-256.

Interpretive Summary: Campylobacters are considered one of the most frequent causes of foodborne infections in developing as well as developed countries. Due to the fastidious nature of Campylobacter and the difficult discrimination between Campylobacter spp., there is a need for methods that have the ability to rapidly identify and differentiate Campylobacter from clinical samples especially in developing countries such as Egypt. In this study, Campylobacter spp. from human and chicken fecal samples were detected by flagellin gene (flaA) Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP) analysis was used for molecular characterization of Campylobacter isolates. Both methods were useful for identification of Campylobacter spp. and discrimination among the human and chicken Campylobacter isolates. Findings from this study indicate that integrating molecular approaches such as flaA PCR-RFLP is beneficial in analyzing Campylobacter isolates from humans and poultry for their similarity and possible relevance for exposure to each other. This knowledge may assist research scientists and epidemiologists in the identification and implementation of appropriate control measures that will reduce the number of cases of human campylobacteriosis associated with the consumption of contaminated poultry and other food products.

Technical Abstract: Campylobacter is a foodborne pathogen which has a potential public health concern worldwide. Due to discriminatory problems encountered by conventional isolation of Campylobacter spp. and its genetic similarities, rapid molecular techniques for its genetic characterization are useful. In this study, Campylobacter spp. from human and chicken fecal samples were detected by flagellin gene (flaA) Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP) was used for molecular characterization of Campylobacter spp. A total of 297 samples consisting of 163 adult human stools (102 from diarrheic patients and 61 from apparently healthy) and 134 chicken feces were subjected to flaA PCR. The clinical strains identified from human and chicken feces and the thirteen Campylobacter reference strains were typed by flaA RFLP using DdeI restriction enzyme. A total of 13 human stool samples (8.27%) and 39 (29.1%) chicken feces samples yielded the genus specific 1.7 Kb amplicon by flaA PCR assay. Reference and fecal strains identified by flaA PCR were differentiated using RFLP analysis into 14 RFLP patterns that categorized into 4 clusters. Eleven human stool isolates had a RFLP genotype identical to the genotype of the reference strain C. jejuni 1997-8. Also, 25 and 10 chicken fecal isolates had fragmentation patterns indistinguishable from those of C. jejuni and C. coli reference strains, respectively. The C. jejuni pattern was the dominant pattern identified from chicken fecal samples and 2 clinical isolates of C. jejuni (Cj7) shared a similarity of 73% with that identified from human stool C. jejuni (Cj3). To the best of our knowledge, this is the first report in Egypt that uses flaA PCR- RFLP as a rapid tool for the detection of Campylobacter spp. directly from human and chicken feces and assessment of genetic similarities among Campylobacter spp.