Undressing of Waddlia chondrophila to enrich its outer membrane proteins to develop a new species‐specific ELISA. Issue 1 (16th January 2014)

• Record Type: Journal Article
• Authors: Lienard, J.; Croxatto, A.; Gervaix, A.; Posfay‐Barbe, K.; Baud, D.; Kebbi‐Beghdadi, C.; Greub, G.
• Is Part Of: New microbes and new infections. Volume 2:Issue 1(2014)
• Page Start: 13
• Page End: 24
• Publication Date: 2014-01-16
• Subjects: Medical microbiology -- Periodicals; Communicable diseases -- Periodicals; Infection -- Periodicals; Microbiology -- Periodicals; Communicable Diseases -- Periodicals; Infection -- Periodicals; 616.9041
• DOI: 10.1002/2052-2975.26
• Languages: English
• ISSNs: 2052-2975

Abstract

<abstract abstract-type="main" id="nmi226-abs-0001"> <title>Abstract</title> <p> <italic>Waddlia chondrophila</italic>, an obligate intracellular bacterium of the Chlamydiales order, is considered as an agent of bovine abortion and a likely cause of miscarriage in humans. Its role in respiratory diseases was questioned after the detection of its DNA in clinical samples taken from patients suffering from pneumonia or bronchiolitis. To better define the role of <italic>Waddlia</italic> in both miscarriage and pneumonia, a tool allowing large‐scale serological investigations of <italic>Waddlia</italic> seropositivity is needed. Therefore, enriched outer membrane proteins of <italic>W. chondrophila</italic> were used as antigens to develop a specific ELISA. After thorough analytical optimization, the ELISA was validated by comparison with micro‐immunofluorescence and it showed a sensitivity above 85% with 100% specificity. The ELISA was subsequently applied to human sera to specify the role of <italic>W. chondrophila</italic> in pneumonia. Overall, 3.6% of children showed antibody reactivity against <italic>W. chondrophila</italic> but no significant difference was observed between children with and without pneumonia. Proteomic analyses were then performed using mass spectrometry, highlighting members of the outer membrane protein family as the dominant proteins. The major <italic>Waddlia</italic> putative immunogenic proteins were identified by immunoblot using positive and negative human sera. The new ELISA represents an efficient tool with high throughput applications. Although no association with pneumonia and <italic>Waddlia</italic> seropositivity was observed, this ELISA could be used to specify the role of <italic>W. chondrophila</italic> in miscarriage and in other diseases.</p> </abstract>