Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice
Abstract Background Rodentibacter (R.) pneumotropicus colonizes the respiratory and urogenital tracts of laboratory mice with a reported moderate serological prevalence from 4 to 13%. Thus, regular tests to identify this pathogen in mice are recommended for animal facilities. However, a recent study...
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doaj-35d27e8e947c448595a6038d3de780222020-11-25T02:11:15ZengBMCBMC Microbiology1471-21802019-02-0119111310.1186/s12866-019-1417-7Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in miceFelix Fingas0Daniela Volke1Rayk Hassert2Juliane Fornefett3Sophie Funk4Christoph Georg Baums5Ralf Hoffmann6Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Universität LeipzigInstitute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Universität LeipzigInstitute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Universität LeipzigInstitute for Bacteriology and Mycology, Faculty of Veterinary Medicine, Universität LeipzigInstitute for Bacteriology and Mycology, Faculty of Veterinary Medicine, Universität LeipzigCenter for Biotechnology and Biomedicine, Universität LeipzigInstitute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Universität LeipzigAbstract Background Rodentibacter (R.) pneumotropicus colonizes the respiratory and urogenital tracts of laboratory mice with a reported moderate serological prevalence from 4 to 13%. Thus, regular tests to identify this pathogen in mice are recommended for animal facilities. However, a recent study indicated that current serological assays are partly insensitive, as C57BL/6 and BALB/c mice infected with R. pneumotropicus were incorrectly screened as seronegative. Results Here, we report a systematic analysis of protein and lipopolysaccharides antigens by immunoblot and ELISA that allowed establishing a sensitive test system able to differentiate between R. pneumotropicus and the closely related species R. heylii. Furthermore, the main immunogen, designated as ‘characteristic antigen for Rodentibacter of laboratory origin 1’ (CARLO-1), was identified by two-dimensional gel electrophoresis followed by immunoblot and tandem mass spectrometry in a preparation of outer membrane proteins. An indirect ELISA relying on the recombinantly expressed protein provided high sensitivity, specificity, and selectivity. The corresponding carlo1 gene was highly conserved (> 97%) among 21 isolates of R. pneumotropicus and R. heylii. Conclusion The newly identified protein CARLO-1 is well suited for the sensitive and specific serological detection of Rodentibacter infections in mice. Indirect differentiation of R. pneumotropicus and R. heylii infections may be possible using an ELISA based on a whole-cell antigen preparation. All four established ELISA systems using a whole-cell preparation, lipopolysaccharides, outer-membrane proteins and protein CARLO-1 as antigen, respectively, outperformed a commercial ELISA in terms of sensitivity.http://link.springer.com/article/10.1186/s12866-019-1417-7ELISACARLO-1Rodentibacter pneumotropicusRodentibacter heyliiPasteurella pneumotropicaFELASA |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Felix Fingas Daniela Volke Rayk Hassert Juliane Fornefett Sophie Funk Christoph Georg Baums Ralf Hoffmann |
spellingShingle |
Felix Fingas Daniela Volke Rayk Hassert Juliane Fornefett Sophie Funk Christoph Georg Baums Ralf Hoffmann Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice BMC Microbiology ELISA CARLO-1 Rodentibacter pneumotropicus Rodentibacter heylii Pasteurella pneumotropica FELASA |
author_facet |
Felix Fingas Daniela Volke Rayk Hassert Juliane Fornefett Sophie Funk Christoph Georg Baums Ralf Hoffmann |
author_sort |
Felix Fingas |
title |
Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice |
title_short |
Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice |
title_full |
Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice |
title_fullStr |
Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice |
title_full_unstemmed |
Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice |
title_sort |
sensitive and immunogen-specific serological detection of rodentibacter pneumotropicus infections in mice |
publisher |
BMC |
series |
BMC Microbiology |
issn |
1471-2180 |
publishDate |
2019-02-01 |
description |
Abstract Background Rodentibacter (R.) pneumotropicus colonizes the respiratory and urogenital tracts of laboratory mice with a reported moderate serological prevalence from 4 to 13%. Thus, regular tests to identify this pathogen in mice are recommended for animal facilities. However, a recent study indicated that current serological assays are partly insensitive, as C57BL/6 and BALB/c mice infected with R. pneumotropicus were incorrectly screened as seronegative. Results Here, we report a systematic analysis of protein and lipopolysaccharides antigens by immunoblot and ELISA that allowed establishing a sensitive test system able to differentiate between R. pneumotropicus and the closely related species R. heylii. Furthermore, the main immunogen, designated as ‘characteristic antigen for Rodentibacter of laboratory origin 1’ (CARLO-1), was identified by two-dimensional gel electrophoresis followed by immunoblot and tandem mass spectrometry in a preparation of outer membrane proteins. An indirect ELISA relying on the recombinantly expressed protein provided high sensitivity, specificity, and selectivity. The corresponding carlo1 gene was highly conserved (> 97%) among 21 isolates of R. pneumotropicus and R. heylii. Conclusion The newly identified protein CARLO-1 is well suited for the sensitive and specific serological detection of Rodentibacter infections in mice. Indirect differentiation of R. pneumotropicus and R. heylii infections may be possible using an ELISA based on a whole-cell antigen preparation. All four established ELISA systems using a whole-cell preparation, lipopolysaccharides, outer-membrane proteins and protein CARLO-1 as antigen, respectively, outperformed a commercial ELISA in terms of sensitivity. |
topic |
ELISA CARLO-1 Rodentibacter pneumotropicus Rodentibacter heylii Pasteurella pneumotropica FELASA |
url |
http://link.springer.com/article/10.1186/s12866-019-1417-7 |
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