Jensen, A.N.; Andersen, M.T.; Dalsgaard, A.; Baggesen, D.L. and Nielsen, E.M. (2005) Development of real-time PCR and hybridization methods for detection and identification of thermophilic Campylobacter spp. in pig faecal samples. Journal of Applied Microbiology, 99, pp. 292-300.
Aims: To develop a real-time (rt) PCR for species differentiation of thermophilic Campylobacter and to develop a method for assessing co-colonization of pigs by Campylobacter spp.
Methods and results: The specificity of a developed 5’nuclease rt-PCR for species-specific identification of C. jejuni, C. coli, C. lari, C. upsaliensis and of a hipO gene nucleotide probe for detection of C. jejuni by colony-blot hybridization were determined by testing a total of 75 reference strains of Campylobacter spp. and related organisms. The rt-PCR method allowed species-specific detection of Campylobacter spp. in naturally infected pig faecal samples after an enrichment step, whereas the hybridization approach enhanced the specific isolation of C. jejuni (present in minority to C. coli) from pigs.
Conclusions: The rt-PCR was specific for Campylobacter jejuni, C. coli, C. lari, and C. upsaliensis and the colony-blot hybridization approach provided an effective tool for isolation of C. jejuni from pig faecal samples typically dominated by C. coli.
Significance and impact of study: Species differentiation between thermophilic Campylobacter is difficult by phenotypic methods and the developed rt-PCR provides an easy and fast method for such differentiation. Detection of C. jejuni by colony hybridization may increase the isolation rate of this species from pig feces.
|EPrint Type:||Journal paper|
|Keywords:||thermophilic Campylobacter, real-time PCR, identification, C. jejuni, hipO gene, hybridization, pigs|
|Subjects:||Animal husbandry > Production systems > Pigs|
|Research affiliation:|| Denmark > DARCOF II (2000-2005) > II.10 (SaCaFree) Bacterial infection risk associated with outdoor organic pig production ...|
Denmark > SOAR - Research School for Organic Agriculture and Food Systems
|Deposited By:||Jensen, Ms Annette Nygaard|
|Deposited On:||17 May 2005|
|Last Modified:||14 Jun 2012 14:28|
|Refereed:||Peer-reviewed and accepted|
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