title: Development and validation of a real-time PCR for Chlamydia suis diagnosis in swine and humans.
abstract: Pigs are the natural host for Chlamydia suis, a pathogen which is phylogenetically highly related to the human pathogen C. trachomatis. Chlamydia suis infections are generally treated with tetracyclines. In 1998, tetracyline resistant C. suis strains emerged on U.S. pig farms and they are currently present in the Belgian, Cypriote, German, Israeli, Italian and Swiss pig industry. Infections with tetracycline resistant C. suis strains are mainly associated with severe reproductive failure leading to marked economical loss. We developed a sensitive and specific TaqMan probe-based C. suis real-time PCR for examining clinical samples of both pigs and humans. The analytical sensitivity of the real-time PCR is 10 rDNA copies/reaction without cross-amplifying DNA of other Chlamydia species. The PCR was successfully validated using conjunctival, pharyngeal and stool samples of slaughterhouse employees, as well as porcine samples from two farms with evidence of reproductive failure and one farm without clinical disease. Chlamydia suis was only detected in diseased pigs and in the eyes of humans. Positive humans had no clinical complaints. PCR results were confirmed by culture in McCoy cells. In addition, Chlamydia suis isolates were also examined by the tet(C) PCR, designed for demonstrating the tetracycline resistance gene tet(C). The tet(C) gene was only present in porcine C. suis isolates.
authors.affiliation: Department of Molecular Biotechnology, Faculty of Bioscience Engineering, University of Ghent, Ghent, Belgium.
authors.forename: Kristien
authors.initials: K
authors.lastname: De Puysseleyr
authors.affiliation: Department of Molecular Biotechnology, Faculty of Bioscience Engineering, University of Ghent, Ghent, Belgium.
authors.forename: Leentje
authors.initials: L
authors.lastname: De Puysseleyr
authors.affiliation: Department of Molecular Biotechnology, Faculty of Bioscience Engineering, University of Ghent, Ghent, Belgium.
authors.forename: Julie
authors.initials: J
authors.lastname: Geldhof
authors.affiliation: Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, University of Ghent, Ghent, Belgium.
authors.forename: Eric
authors.initials: E
authors.lastname: Cox
authors.affiliation: Department of Molecular Biotechnology, Faculty of Bioscience Engineering, University of Ghent, Ghent, Belgium.
authors.forename: Daisy
authors.initials: D
authors.lastname: Vanrompay
mesh_terms: D000818:Animals; D002689:Chlamydia / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D005260:Female; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D013552:Swine
chemical_list: D017931:DNA Primers; D015342:DNA Probes