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Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice
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Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice
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Journal Article
Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice
2019
Overview
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.
Publisher
BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
Subject
/ Antigens
/ Antigens, Bacterial - immunology
/ Bacterial Proteins - immunology
/ Biomedical and Life Sciences
/ CARLO-1
/ ELISA
/ Enzyme-Linked Immunosorbent Assay
/ Enzymes
/ FELASA
/ Female
/ Genes
/ Mice
/ Mycology
/ Pasteurellaceae - immunology
/ Pasteurellaceae - pathogenicity
/ Pasteurellaceae Infections - diagnosis
/ Pasteurellaceae Infections - immunology
/ Pasteurellaceae Infections - veterinary
/ Proteins
/ Rodentibacter pneumotropicus
/ Rodents
/ Virology
