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

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...

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Bibliographic Details
Published in:European journal of clinical microbiology & infectious diseases 2009-12, Vol.28 (12), p.1483-1486
Main Authors: Goy, G., Croxatto, A., Posfay-Barbe, K. M., Gervaix, A., Greub, G.
Format: Article
Language:English
Subjects:
Bacteriological Techniques - methods
Biological and medical sciences
Biomedical and Life Sciences
Biomedicine
Brief Report
Child
Chlamydiales - genetics
Chlamydiales - isolation & purification
DNA Primers - genetics
DNA, Bacterial - genetics
DNA, Ribosomal - genetics
Female
Gram-Negative Bacterial Infections - diagnosis
Gram-Negative Bacterial Infections - microbiology
Humans
Infant
Infant, Newborn
Infectious diseases
Internal Medicine
Medical Microbiology
Medical sciences
Nasopharynx - microbiology
Polymerase Chain Reaction - methods
Reproducibility of Results
Respiratory tract
RNA, Ribosomal, 16S - genetics
Sensitivity and Specificity
Sexually transmitted diseases
STD
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Summary: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.
ISSN:0934-9723
1435-4373
DOI:10.1007/s10096-009-0804-7