An enzyme-linked immunosorbent assay (ELISA) for monitoring rodent colonies for <i>Pasteurella pneumotropica</i> antibodies

Boot, R.; Thuis, H. C. W.; Veenema, J. L.; Bakker, Richard G.

doi:10.1258/002367795781088306

Cited by 17 publications

(6 citation statements)

References 9 publications

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“…The WCA-based ELISA established by our group appears to provide a much higher sensitivity and to differentiate defined positive sera of the two closely related Rodentibacter species. Apparently, the new test improves previously reported WCA-based ELISA protocols [12, 21]. Generally, the reported prevalence may be influenced by insensitive ELISA assays used in routine health monitoring, as shown here for one commercial assay.…”

Section: Discussionsupporting

confidence: 52%

“…Indirect R. pneumotropicus - or R. heylii -specific serological assays, such as enzyme-linked immunosorbent assay (ELISA), have not been described to the best of our knowledge. Currently, applied ELISA rely on inactivated whole cells detecting both P. pneumotropica biotypes Jawetz and Heyl [12, 13]. Manning et al reported a sensitive detection of P. pneumotropica antibodies based on a preparation of uncharacterized cell wall proteins, which were not examined for their specificity to Jawetz and Heyl isolates [14].…”

Section: Introductionmentioning

confidence: 99%

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Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice

et al. 2019

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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. Electronic supplementary material The online version of this article (10.1186/s12866-019-1417-7) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 52%

Section: Introductionmentioning

confidence: 99%

Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice

et al. 2019

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“…The Federation of Laboratory Animal Science Association (FELASA) lists P. pneumotropica as an important pathogen in mice, rats and hamsters and recommends the examination every 3 months [ 16 ]. Various monitoring methods are described including PCR [ 4 , 5 , 17 ] and indirect enzyme-linked immunosorbent assays (ELISAs) [ 18 – 20 ].…”

Section: Introductionmentioning

confidence: 99%

Comparative analysis of humoral immune responses and pathologies of BALB/c and C57BL/6 wildtype mice experimentally infected with a highly virulent Rodentibacter pneumotropicus (Pasteurella pneumotropica) strain

et al. 2018

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BackgroundMice are a natural host for Rodentibacter (R.) pneumotropicus. Despite specific monitoring, it is still one of the most important infectious agents in laboratory animals. The objective of this study was to determine the virulence of a prevalent pathotype of R. pneumotropicus and characterize the host response in a new animal model.ResultsIntranasal infection of C57BL/6 and BALB/c mice with a R. pneumotropicus strain (JF4Ni) bearing the genes of the three known repeats in toxin (RTX) toxins resulted in an unprecedented high mortality and morbidity above 50 and 80%, respectively. Morbidity was associated with severe weight loss as well as conjunctivitis and dyspnea. A main pathology was a catarrhal purulent to necrotic bronchopneumonia. Specific immune globuline (Ig) A was detected in tracheonasal lavages of most surviving mice which were still colonized by R. pneumotropicus. Furthermore, all surviving animals showed a distinct production of IgG antibodies. To differentiate T-helper cell (Th) 1 and Th2 immune responses we used subclasses of IgGs as indicators. Mean ratios of IgG2b to IgG1 were below 0.8 in sera drawn from both mice strains prior infection and from BALB/c mice post infection. In contrast, C57BL/6 mice had a mean IgG2b/IgG1 ratio of 1.6 post infection indicating a Th1 immune response in C57BL/6 versus a Th2 response in BALB/c mice associated with a tenfold higher bacterial load in the lung. In accordance with a Th1 response high antigen-specific IgG2c titers were detected in the majority of surviving C57BL/6 mice.ConclusionsR. pneumotropicus JF4Ni is a highly virulent strain causing severe pneumonia and septicemia after intranasal infection of C57BL/6 and BALB/c mice. Persisting infections in the two mice strains are associated with Th1 and Th2 immune responses, respectively, and differences in the bacterial burden of the lung. The described model is ideally suited for future vaccination studies using the natural host.Electronic supplementary materialThe online version of this article (10.1186/s12866-018-1186-8) contains supplementary material, which is available to authorized users.

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“…Nonetheless, serology has been employed, along with other diagnostic methodologies, to monitor laboratory animals for infections with fastidious bacteria such as Clostridium piliforme (Riley et al, 1994;Waggie et al, 1987), Helicobacter hepaticus , and CAR bacillus (Lukas et al, 1987b;Matsushita et aL, 1987). Serologic assays have also been developed for gram-negative bacteria such as Pasteurella pneumotropica (Boot et al, 1995), P. multocida (Lukas et al, 1987a), and Bordetella bronchiseptica from which specific lipopolysaccharide antigen can be purified (Manning et al, 1987).…”

Section: Serology For Detection Of Antibodies To Infectious Agentsmentioning

confidence: 99%

Microbiological Quality Control for Laboratory Rodents and Lagomorphs

Shek

Smith

Pritchett-Corning

2015

Laboratory Animal Medicine

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An enzyme-linked immunosorbent assay (ELISA) for monitoring rodent colonies for Pasteurella pneumotropica antibodies

Cited by 17 publications

References 9 publications

Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice

Sensitive and immunogen-specific serological detection of Rodentibacter pneumotropicus infections in mice

Comparative analysis of humoral immune responses and pathologies of BALB/c and C57BL/6 wildtype mice experimentally infected with a highly virulent Rodentibacter pneumotropicus (Pasteurella pneumotropica) strain

Microbiological Quality Control for Laboratory Rodents and Lagomorphs

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