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Development and validation of real-time PCR for Chlamydia suis diagnosis in swine and humans

Kristien De Puysseleyr (UGent) , Leentje De Puysseleyr (UGent) , Julie Geldhof (UGent) , Eric Cox (UGent) and Daisy Vanrompay (UGent)
(2014) PLOS ONE. 9(5).
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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.
Keywords
ASSAY, SENSITIVITY, PSITTACI, IDENTIFICATION, PNEUMONIAE, INFECTION, PIGS, CHLAMYDOPHILA-FELIS, QUANTITATIVE PCR, REPRODUCTIVE FAILURE

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Chicago
De Puysseleyr, Kristien, Leentje De Puysseleyr, Julie Geldhof, Eric Cox, and Daisy Vanrompay. 2014. “Development and Validation of Real-time PCR for Chlamydia Suis Diagnosis in Swine and Humans.” Plos One 9 (5).
APA
De Puysseleyr, K., De Puysseleyr, L., Geldhof, J., Cox, E., & Vanrompay, D. (2014). Development and validation of real-time PCR for Chlamydia suis diagnosis in swine and humans. PLOS ONE, 9(5).
Vancouver
1.
De Puysseleyr K, De Puysseleyr L, Geldhof J, Cox E, Vanrompay D. Development and validation of real-time PCR for Chlamydia suis diagnosis in swine and humans. PLOS ONE. 2014;9(5).
MLA
De Puysseleyr, Kristien et al. “Development and Validation of Real-time PCR for Chlamydia Suis Diagnosis in Swine and Humans.” PLOS ONE 9.5 (2014): n. pag. Print.
@article{4427984,
  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.},
  articleno    = {e96704},
  author       = {De Puysseleyr, Kristien and De Puysseleyr, Leentje and Geldhof, Julie and Cox, Eric and Vanrompay, Daisy},
  issn         = {1932-6203},
  journal      = {PLOS ONE},
  keywords     = {ASSAY,SENSITIVITY,PSITTACI,IDENTIFICATION,PNEUMONIAE,INFECTION,PIGS,CHLAMYDOPHILA-FELIS,QUANTITATIVE PCR,REPRODUCTIVE FAILURE},
  language     = {eng},
  number       = {5},
  pages        = {7},
  title        = {Development and validation of real-time PCR for Chlamydia suis diagnosis in swine and humans},
  url          = {http://dx.doi.org/10.1371/journal.pone.0096704},
  volume       = {9},
  year         = {2014},
}

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