Screening and Rapid Identification of Campylobacter Spp. DNA by FlaA PCR Based Method on Chicken and Human Fecal Samples in Egypt
DOI:
https://doi.org/10.3923/ijps.2015.252.256Keywords:
Campylobacter, flaA PCR, human stool, poultry fecesAbstract
Due to culturability requirements encountered by the conventional isolation of Campylobacter spp., rapid molecular techniques for its direct identification from clinical samples are useful. In this study, Campylobacter spp. DNA from human stool and chicken fecal samples were detected by flagellin gene (flaA) PCR. A total of 297 samples consisting of 163 adult human stools (102 from diarrheic patients and 61 from healthy persons) and 134 chicken feces were subjected to flaA PCR. Ten reference strains of Campylobacter spp. were included in this study as positive controls. Thirteen stool samples (7.98%) from the human fecal samples and 39 chicken fecal samples (29.1%) yielded the genus specific 1.7 Kb amplicon of Campylobacter spp. Eight (7.84%) diarrheic human stool specimens out of 102 samples and 5 (8.2%) apparently healthy human stool specimens out of 61 samples were positive by flaA PCR assay. All the Campyloacter reference strains examined giving the specific amplicon of 1.7 Kb. The existence of Campylobacter spp. DNA detected by flaA PCR in poultry and human samples taken from locations of Egypt highlights the zoonotic potential of Campylobacter. To the best of our knowledge, this is the first report in Egypt that uses flaA PCR as a rapid screening method for the direct detection of Campylobacter spp. from human and chicken feces.
References
Al Amri, A., A.C. Senok, A.Y. Ismaeel, A.E. Al-Mahmeed and G.A. Botta, 2007. Multiplex PCR for direct identification of Campylobacter spp. in human and chicken stools. J. Med. Microbiol., 56: 1350-1355.
Allos, B.M., 2001. Campylobacter jejuni infections: Update on emerging issues and trends. Clin. Infect. Dis., 32: 1201-1206.
Coker, A.O., R.D. Isokpehi, B.N. Thomas, K.O. Amisu and C.L. Obi, 2002. Human campylobacteriosis in developing countries. Emerg. Infect. Dis., 8: 237-244.
Corry, J.E. and H.I. Atabay, 2001. Poultry as a source of Campylobacter and related organisms. Symp. Ser. Soc. Applied Microbiol., 30: 96S-114S.
Endtz, H.P., G.J. Ruijs, A.H. Zwinderman, T. van der Reijden, M. Biever and R.P. Mouton, 1991. Comparison of six media, including a semisolid agar, for the isolation of various Campylobacter species from stool specimens. J. Clin. Microbiol., 29: 1007-1010.
Fitzgerald, C., K. Stanley, S. Andrew and K. Jones, 2001. Use of pulsed-field gel electrophoresis and flagellin gene typing in identifying clonal groups of Campylobacter jejuni and Campylobacter coli in farm and clinical environments. Applied Environ. Microbiol., 67: 1429-1436.
Fode-Vaughan, K.A., C.F. Wimpee, C.C. Remsen and M.L.P. Collins, 2001. Detection of bacteria in environmental samples by direct PCR without DNA extraction. Biotechniques, 31: 598-607.
Guyard-Nicodeme, M., O. Tresse, E. Houard, F. Jugiau and C. Courtillon et al., 2013. Characterization of Campylobacter spp. transferred from naturally contaminated chicken legs to cooked chicken slices via a cutting board. Int. J. Food Microbiol., 164: 7-14.
Hinton, Jr.A., 2006. Comparison of growth of campylobacteriaceae on media supplemented with organic acids and on commercially available media. Int. J. Poult. Sci., 5: 99-103.
Hinton, Jr.A., J.A. Cason, M.E. Hume and K.D. Ingram, 2004. Use of MIDI-fatty acid methyl ester analysis to monitor the transmission of Campylobacter during commercial poultry processing. J. Food Prot., 67: 1610-1616.
Inglis, G.D. and L.D. Kalischuk, 2003. Use of PCR for direct detection of Campylobacter species in bovine feces. Applied Environ. Microbiol., 69: 3435-3447.
LaGier, M.L., L.A. Joseph, T.V. Passaretti, K.A. Musser and N.M. Cirino, 2004. A real-time multiplexed PCR assay for rapid detection and differentiation of Campylobacter jejuni and Campylobacter coli. Molecul. Cell. Prob., 18: 275-282.
Lawson, A.J., D. Linton and J. Stanley, 1998. 16S rRNA gene sequences of Candidatus Campylobacter hominis, a novel uncultivated species, are found in the gastrointestinal tract of healthy humans. Microbiology, 144: 2063-2071.
Linton, D., A.J. Lawson, R.J. Owen and J. Stanley, 1997. PCR detection, identification to species level and fingerprinting of Campylobacter jejuni and Campylobacter coli direct from diarrheic samples. J. Clin. Microbiol., 35: 2568-2572.
McOrist, A.L., M. Jackson and A.R. Bird, 2002. A comparison of five methods for extraction of bacterial DNA from human faecal samples. J. Microbiol. Methods, 50: 131-139.
Nachamkin, I., K. Bohachick and C.M. Patton, 1993. Flagellin gene typing of Campylobacter jejuni by restriction fragment length polymorphism analysis. J. Clin. Microbiol., 31: 1531-1536.
Oosterom, J., S. Notermans, H. Karman and G.B. Engels, 1983. Origin and prevalence of Campylobacter jejuni in poultry processing. J. Food Prot., 46: 339-344.
Parkar, S.F.D., D. Sachdev, N. de Souza, A. Kamble and G. Suresh et al., 2013. Prevalence, seasonality and antibiotic susceptibility of thermophilic Campylobacters in ceca and carcasses of poultry birds in the live-bird market. Afr. J. Microbiol. Res., 7: 2442-2453.
Persson, S. and K.E.P. Olsen, 2005. Multiplex PCR for identification of campylobacter coli and Campylobacter jejuni from pure cultures and directly on stool samples. J. Med. Microbiol., 54: 1043-1047.
USDA., 2001. Egypt poultry and products. Annual GAIN Report No. EG1020, U.S. Department of Agriculture, Foreign Agricultural Service, Washington, DC., USA.
Wilson, I.G., 1997. Inhibition and facilitation of nucleic acid amplification. Applied Environ. Microbiol., 63: 3741-3751.
Downloads
Published
Issue
Section
License
Copyright (c) 2015 Asian Network for Scientific Information
This work is licensed under a Creative Commons Attribution 4.0 International License.
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited.
