Monoclonal antibodies against all known variants of EspA: development of a simple diagnostic test for enteropathogenic Escherichia coli based on a key virulence factor.

Autor:	Praekelt U; Department of Genetics, University of Leicester, Leicester, UK., Reissbrodt R; Abteilung für Infektionskrankheiten, Robert Koch Institut, Wernigerode, Germany., Kresse A; Abteilung für Infektionskrankheiten, Robert Koch Institut, Wernigerode, Germany., Pavankumar A; Centre for Biotechnology, Anna University, Chennai, India., Sankaran K; Centre for Biotechnology, Anna University, Chennai, India., James R; Department of Infection, Immunity and Inflammation, University of Leicester, Leicester, UK., Jesudason M; Christian Medical College, Vellore, India., Anandan S; Christian Medical College, Vellore, India., Prakasam A; Christian Medical College, Vellore, India., Balaji V; Christian Medical College, Vellore, India., Dutta S; National Institute of Cholera and Enteric Diseases, Kolkata, India., Dutta S; National Institute of Cholera and Enteric Diseases, Kolkata, India., Ramamurthy T; National Institute of Cholera and Enteric Diseases, Kolkata, India., Fischer R; R-Biopharm AG, Darmstadt, Germany., Sander P; R-Biopharm AG, Darmstadt, Germany., Schaumann R; Institut für Medizinische Mikrobiologie, Universität Leipzig, Leipzig, Germany., Navarro A; Department of Public Health, Faculty of Medicine, National Autonomous University of Mexico, Mexico City, Mexico., Williams P; Department of Genetics, University of Leicester, Leicester, UK.
Jazyk:	angličtina
Zdroj:	Journal of medical microbiology [J Med Microbiol] 2014 Dec; Vol. 63 (Pt 12), pp. 1595-1607. Date of Electronic Publication: 2014 Sep 17.
DOI:	10.1099/jmm.0.076323-0
Abstrakt:	Enteropathogenic Escherichia coli (EPEC) are a major cause of infant diarrhoea in developing countries and a significant public health issue in industrialized countries. Currently there are no simple tests available for the diagnosis of EPEC. Serology of O-antigens is widely used routinely in many laboratories throughout the world, even though it has been known for many years to be an unreliable indicator of EPEC virulence. We have developed a simple, low-cost immunodiagnostic test based on the EspA filament, an essential virulence factor of EPEC and the related enterohaemorrhagic E. coli (EHEC). Using recombinant proteins of the five major variants of EspA as immunogens, we raised a panel of three monoclonal antibodies in mice that detects all variants of the native target in bacterial cultures. The antibodies proved suitable for application in sandwich-type assays, including ELISA and lateral flow immunoassays (LFI). Prototypes for both assays were specific for EPEC and EHEC strains when tested against a panel of control micro-organisms. We have also developed a simple, affordable culture medium, A/E medium, which optimizes expression of EspA allowing improved sensitivity of detection compared with standard Dulbecco's modified Eagle's medium. Together these reagents form the basis of robust, informative tests for EPEC for use especially in developing countries but also for routine screening in any clinical laboratory. (© 2014 The Authors.)
Databáze:	MEDLINE
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