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          Institute: MPI für molekulare Genetik     Collection: Department of Vertebrate Genomics     Display Documents



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ID: 275619.0, MPI für molekulare Genetik / Department of Vertebrate Genomics
Bacterial protein microarrays for identification of new potential diagnostic markers for Neisseria meningitidis infections
Authors:Steller, Sigrid; Angenendt, Philipp; Cahill, Dolores J.; Heuberger, Sigrid; Lehrach, Hans; Kreutzberger, Jürgen
Language:English
Date of Publication (YYYY-MM-DD):2005-04-25
Title of Journal:PROTEOMICS
Volume:5
Issue / Number:8
Start Page:2048
End Page:2055
Copyright:© 2005 WILEY-VCH Verlag GmbH & Co. KGaA
Review Status:not specified
Audience:Experts Only
Abstract / Description:Neisseria meningitidis is the most common cause of meningitis and causes epidemic outbreaks. One trait of N. meningitidis, which is associated with most of the currently recognized virulence determinants, is the presence of phase-variable genes that are suspected to enhance its ability to cause an invasive disease. To detect the immune responses to phase-variable expressed proteins, we applied protein microarray technology for the screening of meningitis patient sera. We amplified all 102 known phase-variable genes from N. meningitidis serogroup B strain MC58 by polymerase chain reaction and subcloned them for expression in Escherichia coli. With this approach, we were able to express and purify 67 recombinant proteins representing 66% of the annotated genes. These were spotted robotically onto coated glass slides to generate protein microarrays, which were screened using 20 sera of patients suffering from meningitis, as well as healthy controls. From these screening experiments, 47 proteins emerged as immunogenic, exhibiting a variable degree of seroreactivity with some of the patient sera. Nine proteins elicited an immune response in more than three patients, with one of them, the phase-variable opacity protein OpaV (NMB0442), showing responses in 11 patient sera. This is the first time that protein microarray technology has been applied for the investigation of genetic phase variation in pathogens. The identification of disease-specific proteins is a significant target in biomedical research, as such proteins may have medical, diagnostic, and commercial potential as disease markers.
Free Keywords:Opacity proteins; Pathogen; Phase variation; Protein chip; Vaccine
Comment of the Author/Creator:Email: Jürgen Kreutzberger (kreutzbg@molgen.mpg.de)
External Publication Status:published
Document Type:Article
Version Comment:Automatic journal name synchronization
Communicated by:Hans Lehrach
Affiliations:MPI für molekulare Genetik
External Affiliations:Ruhr-University Bochum, Medical Proteome Center, Bochum, Germany;
PROTAGEN AG, Dortmund, Germany;
The Centre for Human Proteomics, Royal College of Surgeons in Ireland, Dublin, Ireland;
Nationale Referenzzentrale für Meningokokken, AGES BSU-Graz, Graz, Austria
Identifiers:ISSN:1615-9853
DOI:10.1002/pmic.200401097
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