Antigenicity of Recombinant Maltose Binding Protein-Mycobacterium avium subsp. paratuberculosis Fusion Proteins with and without Factor Xa Cleaving

Gurung, Ratna B.; Begg, Douglas J.; Purdie, Auriol C.; Bannantine, John P.; Whittington, Richard J.

doi:10.1128/cvi.00596-13

Cited by 9 publications

(6 citation statements)

References 47 publications

(71 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…MAP3531, a secreted fibronectin binding protein (Antigen 85C) has also been evaluated in several previous studies where reactivity in the infected group was higher but not significantly different from the control 22 , however other studies have found it to be significantly (P < 0.05) more reactive with ELISA 21 or immunoblotting 29 based assays. Other candidate proteins such as MAP2705 and MAP2764 that were not identified as significantly reactive antigens in the current investigation, were also previously found to not differ in reactivity between groups in previous studies 29,30 .…”

Section: Discussionsupporting

confidence: 70%

Identification of Sero-Diagnostic Antigens for the Early Diagnosis of Johne’s Disease using MAP Protein Microarrays

Bannantine

Campo

Randall

et al. 2019

Sci Rep

Self Cite

View full text Add to dashboard Cite

Considerable effort has been directed toward controlling Johne’s disease (JD), a chronic granulomatous intestinal inflammatory disease caused by Mycobacterium avium subsp. paratuberculosis (MAP) in cattle and other ruminants. However, progress in controlling the spread of MAP infection has been impeded by the lack of reliable diagnostic tests that can identify animals early in the infection process and help break the transmission chain. To identify reliable antigens for early diagnosis of MAP infection, we constructed a MAP protein array with 868 purified recombinant MAP proteins, and screened a total of 180 well-characterized serum samples from cows assigned to 4 groups based on previous serological and fecal test results: negative low exposure (NL, n = 30); negative high exposure (NH, n = 30); fecal-positive, ELISA-negative (F + E−, n = 60); and both fecal- and ELISA-positive (F + E+, n = 60). The analyses identified a total of 49 candidate antigens in the NH, F + E−, and F + E+ with reactivity compared with the NL group (p < 0.01), a majority of which have not been previously identified. While some of the antigens were identified as reactive in only one of the groups, others showed reactivity in multiple groups, including NH (n = 28), F + E− (n = 26), and F + E+ (n = 17) groups. Using combinations of top reactive antigens in each group, the results reveal sensitivities of 60.0%, 73.3%, and 81.7% in the NH, F + E−, and F + E+, respectively at 90% specificity, suggesting that early detection of infection in animals may be possible and enable better opportunities to reduce within herd transmission that may be otherwise missed by traditional serological assays that are biased towards more heavily infected animals. Together, the results suggest that several of the novel candidate antigens identified in this study, particularly those that were reactive in the NH and F + E− groups, have potential utility for the early sero-diagnosis of MAP infection.

show abstract

Section: Discussionsupporting

confidence: 70%

Identification of Sero-Diagnostic Antigens for the Early Diagnosis of Johne’s Disease using MAP Protein Microarrays

Bannantine

Campo

Randall

et al. 2019

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…The ortholog MAP1569 (ModD) was also identified as a secreted protein that was recognized by sera collected from naturally infected cows [ 22 , 23 ]. The ortholog MAP0834c, a two component system transcriptional regulator, was recognized by sera from naturally MAP infected sheep as a significantly reactive antigen [ 24 ]. Another ortholog MAP1272c, an invasion-associated protein, has been identified in several studies as one a promising antigen [ 24 , 25 ] and recently further characterized on crystal structures, combined with functional assays [ 26 ].…”

Section: Resultsmentioning

confidence: 99%

“…The ortholog MAP0834c, a two component system transcriptional regulator, was recognized by sera from naturally MAP infected sheep as a significantly reactive antigen [ 24 ]. Another ortholog MAP1272c, an invasion-associated protein, has been identified in several studies as one a promising antigen [ 24 , 25 ] and recently further characterized on crystal structures, combined with functional assays [ 26 ]. The ortholog MAP0900 (P35), a conserved membrane protein, was recognized by 100% of animals including cattle, goats and sheep with Johne’s disease in the clinical stage and 75% of cattle in the sub-clinical stage [ 27 ], as well as 75% of patients with Crohn’s disease [ 28 ].…”

Section: Resultsmentioning

confidence: 99%

Identification of sero-reactive antigens for the early diagnosis of Johne’s disease in cattle

Bannantine

Campo

et al. 2017

PLoS ONE

Self Cite

View full text Add to dashboard Cite

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease (JD), a chronic intestinal inflammatory disease of cattle and other ruminants. JD has a high herd prevalence and causes serious animal health problems and significant economic loss in domesticated ruminants throughout the world. Since serological detection of MAP infected animals during the early stages of infection remains challenging due to the low sensitivity of extant assays, we screened 180 well-characterized serum samples using a whole proteome microarray from Mycobacterium tuberculosis (MTB), a close relative of MAP. Based on extensive testing of serum and milk samples, fecal culture and qPCR for direct detection of MAP, the samples were previously assigned to one of 4 groups: negative low exposure (n = 30, NL); negative high exposure (n = 30, NH); fecal positive, ELISA negative (n = 60, F+E-); and fecal positive, ELISA positive (n = 60, F+E+). Of the 740 reactive proteins, several antigens were serologically recognized early but not late in infection, suggesting a complex and dynamic evolution of the MAP humoral immune response during disease progression. Ordinal logistic regression models identified a subset of 47 candidate proteins with significantly different normalized intensity values (p<0.05), including 12 in the NH and 23 in F+E- groups, suggesting potential utility for the early detection of MAP infected animals. Next, the diagnostic utility of four MAP orthologs (MAP1569, MAP2942c, MAP2609, and MAP1272c) was assessed and reveal moderate to high diagnostic sensitivities (range 48.3% to 76.7%) and specificity (range 96.7% to 100%), with a combined 88.3% sensitivity and 96.7% specificity. Taken together, the results of our analyses have identified several candidate MAP proteins of potential utility for the early detection of MAP infection, as well individual MAP proteins that may serve as the foundation for the next generation of well-defined serological diagnosis of JD in cattle.

show abstract

“…These findings demonstrated that the specificity of the indirect ELISA method was better than that of PAT. However, MBP fusion protein, which was used as the coating antigen could also react with antibodies against MBP, and such antibodies have been found to have minor seroreactivity [18]. MBP-tagged avian influenza M2 protein has been developed as a coating antigen in the M2e-MBP ELISA method for differentiating avian influenza infected chickens from vaccinated ones [19].…”

Section: Discussionmentioning

confidence: 99%

Purification of recombinant IpaJ to develop an indirect ELISA-based method for detecting Salmonella enterica serovar Pullorum infections in chickens

Yue

Yin

et al. 2019

BMC Vet Res

View full text Add to dashboard Cite

BackgroundSalmonella enterica serovar Pullorum is a host-restricted serotype causing infection in poultry. The pathogen can not only cause acute infection in young chicks with high mortality and morbidity, but also persist in adult chickens without evident clinical symptoms and lead to vertical transmission. To eradicate S. Pullorum in poultry farms, it is necessary to establish an efficient method to monitor the prevalence of the pathogen in adult chickens. The protein IpaJ is a specific immunogen in S. Pullorum and is not detected in closely related serotypes, such as S. Gallinarum and S. Enteritidis.ResultsIn the present study, IpaJ was expressed as a recombinant fusion protein MBP-IpaJ in E. coli. The purified MBP-IpaJ was used as a coating antigen to develop an indirect ELISA assay, which was applied to the detection of S. Pullorum infection in chickens. The indirect ELISA assay demonstrated that antibodies produced against IpaJ were detectable in antisera of chickens infected with S. Pullorum in the second week, stably increased until the tenth week, and persisted at a high level in the following two weeks. Furthermore, the ELISA method detected four positive samples out of 200 clinical antiserum samples collected from a poultry farm, and the positive samples were confirmed to be reacted with S. Pullorum using the standard plate agglutination test.ConclusionsThe established indirect ELISA using the IpaJ protein is a novel method for specific detection of S. Pullorum infection, and contribute to eradication of pullorum disease in the poultry industry.Electronic supplementary materialThe online version of this article (10.1186/s12917-018-1753-0) contains supplementary material, which is available to authorized users.

show abstract

Antigenicity of Recombinant Maltose Binding Protein-Mycobacterium avium subsp. paratuberculosis Fusion Proteins with and without Factor Xa Cleaving

Cited by 9 publications

References 47 publications

Identification of Sero-Diagnostic Antigens for the Early Diagnosis of Johne’s Disease using MAP Protein Microarrays

Identification of Sero-Diagnostic Antigens for the Early Diagnosis of Johne’s Disease using MAP Protein Microarrays

Identification of sero-reactive antigens for the early diagnosis of Johne’s disease in cattle

Purification of recombinant IpaJ to develop an indirect ELISA-based method for detecting Salmonella enterica serovar Pullorum infections in chickens

Contact Info

Product

Resources

About