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Development of a real-time PCR for the specific detection of Waddlia chondrophila in clinical samples

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Abstract

Waddlia chondrophila is considered as an emerging human pathogen likely involved in miscarriage and lower respiratory tract infections. Given the low sensitivity of cell culture to recover such an obligate intracellular bacteria, molecular-based diagnostic approaches are warranted. We thus developed a real-time PCR that amplifies Waddlia chondrophila DNA. Specific primers and probe were selected to target the 16S rRNA gene. The PCR specifically amplified W. chondrophila but did not amplify other related-bacteria such as Parachlamydia acanthamoebae, Simkania negevensis and Chlamydia pneumoniae. The PCR exhibited a good intra-run and inter-run reproducibility and a sensitivity of less than ten copies of the positive control. This real-time PCR was then applied to 32 nasopharyngeal aspirates taken from children with bronchiolitis not due to respiratory syncytial virus (RSV). Three samples revealed to be Waddlia positive, suggesting a possible role of this Chlamydia-related bacteria in this setting.

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Acknowledgements

We thank Sébastien Aeby and Nicola Casson for technical help. This project was funded by Swiss National Science Foundation (SNSF) grant number 3200BO-116445. Gilbert Greub is supported by the Leenards Foundation through a career award entitled “Bourse Leenards pour la relève académique en médecine clinique à Lausanne”.

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Authors and Affiliations

  1. Center for Research on Intracellular Bacteria, Institute of Microbiology, University Hospital Center and University of Lausanne, 1011, Lausanne, Switzerland

    G. Goy, A. Croxatto & G. Greub

  2. Children’s Hospital of Geneva, University Hospitals of Geneva, Geneva, Switzerland

    K. M. Posfay-Barbe & A. Gervaix

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  1. G. Goy
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  2. A. Croxatto
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  3. K. M. Posfay-Barbe
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  4. A. Gervaix
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  5. G. Greub
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Corresponding author

Correspondence to G. Greub.

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Goy, G., Croxatto, A., Posfay-Barbe, K.M. et al. Development of a real-time PCR for the specific detection of Waddlia chondrophila in clinical samples. Eur J Clin Microbiol Infect Dis 28, 1483–1486 (2009). https://doi.org/10.1007/s10096-009-0804-7

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  • DOI: https://doi.org/10.1007/s10096-009-0804-7

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