scholarly journals Improved Skin Test for Differential Diagnosis of Bovine Tuberculosis by the Addition of Rv3020c-Derived Peptides

2012 ◽  
Vol 19 (4) ◽  
pp. 620-622 ◽  
Author(s):  
Gareth J. Jones ◽  
Adam Whelan ◽  
Derek Clifford ◽  
Mick Coad ◽  
H. Martin Vordermeier

ABSTRACTA peptide cocktail derived from the mycobacterial antigens ESAT-6, CFP-10, and Rv3615c allowed differentiation betweenMycobacterium bovis-infected and M. bovis bacillus Calmette-Guérin (BCG)-vaccinated cattle when used as a skin test reagent for a “DIVA” test (i.e., a test capable ofdifferentiatinginfected and uninfectedvaccinatedanimals). Addition of the antigen Rv3020c improves the diagnostic sensitivity without compromising specificity in the face of BCG or Johne's disease vaccination.

Author(s):  
Berecha Bayissa ◽  
Aboma Zewude ◽  
Adane Worku ◽  
Balako Gumi ◽  
Stefan Berg ◽  
...  

Bovine tuberculosis (bTB) challenges intensive dairy production in Ethiopia and implementation of the test and slaughter control strategy is not economically acceptable in the country. Vaccination of cattle with Bacillus Calmette-Guerin (BCG) could be an important adjunct to control, which would require a diagnostic test to differentiate Mycobacterium bovis (M. bovis)-infected and BCG-vaccinated animals (DIVA role). This study describes evaluation of a DIVA skin test (DST) that is based on a cocktail (DSTc) or fusion (DSTf) of specific (ESAT-6, CFP-10 and Rv3615c) M. bovis proteins in Zebu-Holstein crossbred cattle in Ethiopia. The study animals used were 74 calves (35 BCG-vaccinated and 39 unvaccinated) aged less than three weeks at the start and 68 known bTB positive cows. Six weeks after vaccination, the 74 calves were tested with DSTc and the single intradermal cervical comparative tuberculin (SICCT) test. The cows were tested with DSTc and SICCT test. Reactions to DSTc were not observed in BCG-vaccinated and unvaccinated calves while SICCT test reactions were detected in vaccinated calves. DSTc reactions were detected in 95.6% of the cows and single intradermal tuberculin (SIT) positive reactions were found in 98.2% (95% confidence interval, CI, 92.1–100%). The sensitivity of DSTc was 95.6% (95% CI, 87.6–99.1%), and significantly (P<0.001) higher than the sensitivity (75%, 95% CI, 63.0-84.7%) of the SICCT test at 4mm cutoff. DSTf and DSTc reactions were correlated (r = 0.75; 95% CI =0.53–0.88). In conclusion, DSTc could differentiate M. bovis-infected from BCG-vaccinated cattle in Ethiopia. DST had higher sensitivity than the SICCT test. Hence, DSTc could be used as a diagnostic tool for bTB if BCG vaccination is implemented for the control of bTB in Ethiopia and other countries.


2013 ◽  
Vol 20 (10) ◽  
pp. 1559-1562 ◽  
Author(s):  
Bryce M. Buddle ◽  
R. Glyn Hewinson ◽  
H. Martin Vordermeier ◽  
D. Neil Wedlock

ABSTRACTVaccination of cattle with a commercial human tuberculosis (TB) vaccine,Mycobacterium bovisbacillus Calmette-Guérin (BCG) Danish, at a dose equivalent to 5 human doses of BCG has protected these animals against TB in field and experimental trials. There is interest in determining whether a 10-fold-lower dose could still protect cattle but not induce a tuberculin intradermal test response. Two groups of calves (n= 9/group) were vaccinated subcutaneously with a lyophilized BCG Danish vaccine containing either 0.5 (1 × 105to 4 × 105CFU) or 5 (1 × 106to 4 × 106CFU) human doses of BCG Danish, with an additional group of 10 calves serving as nonvaccinated controls. Fifteen weeks after vaccination, these animals were challenged intratracheally with 5 × 103CFU of virulentM. bovisand another 15 weeks later were slaughtered and examined for the presence of tuberculous lesions. Vaccination of the calves with either 0.5 or 5 equivalent human doses of BCG Danish induced similar levels of protection against challenge withM. bovis, with both groups showing significant reductions in the pathological and microbiological parameters compared to those for the the control group (P< 0.05). Vaccination with either of the two BCG doses induced similar numbers of animals responding to the tuberculin intradermal test at 11 weeks postvaccination. Vaccination with a 0.5 equivalent human dose of a commercial lyophilized BCG vaccine can protect cattle against challenge withM. bovis.


2017 ◽  
Vol 24 (5) ◽  
Author(s):  
Gareth J. Jones ◽  
Mick Coad ◽  
Bhagwati Khatri ◽  
Javier Bezos ◽  
Natalie A. Parlane ◽  
...  

ABSTRACT Mycobacterium bovis BCG vaccination sensitizes cattle to bovine tuberculin, which compromises the use of the current bovine tuberculosis (TB) surveillance tests. Although the performance of a blood test (that utilizes antigens expressed by Mycobacterium bovis but not by BCG) capable of discriminating infected from vaccinated animals (DIVA interferon gamma test [DIT]) has been evaluated in naturally infected TB field reactors, there is a need to perform similar analysis in a BCG-vaccinated M. bovis-infected population. Furthermore, we explored different scenarios under which a DIT may be implemented alongside BCG vaccination: (i) serial testing to resolve potential false-positive skin test results or (ii) a standalone test to replace the single intradermal comparative cervical tuberculin (SICCT) skin test. Our results demonstrated significantly better relative test sensitivity when the DIT was evaluated in a serial test scenario. Direct comparison of pre- and post-skin test blood samples revealed that the SICCT test induced significant boosting of the gamma interferon response in M. bovis-infected animals to both the ESAT-6–CFP-10 and Rv3615c peptide cocktails that comprise the DIT, which persisted for the ESAT-6–CFP-10 reagent for at least 14 days. Importantly, no similar boosting effects were observed in noninfected BCG vaccinates, suggesting that DIVA blood testing after a recent skin test would have minimal impact on test specificity.


2012 ◽  
Vol 19 (3) ◽  
pp. 346-351 ◽  
Author(s):  
W. R. Waters ◽  
T. C. Thacker ◽  
B. J. Nonnecke ◽  
M. V. Palmer ◽  
I. Schiller ◽  
...  

ABSTRACTGamma interferon (IFN-γ)-induced protein 10 (IP-10) has recently shown promise as a diagnostic biomarker ofMycobacterium tuberculosisinfection of humans. The aim of the current study was to compare IP-10 and IFN-γ responses uponMycobacterium bovisinfection in cattle by using archived samples from two aerosol inoculation studies. In the first study (104CFUM. bovisby aerosol,n= 7),M. bovispurified protein derivative (PPDb)-specific IP-10 and IFN-γ gene expression was detected as early as 29 days after challenge. PPDb-specific IP-10 and IFN-γ mRNA responses followed a similar pattern of expression over the course of this study and were highly correlated (r= 0.87). In the second study (105CFUM. bovisby aerosol,n= 5), IP-10 and IFN-γ (protein) responses to mycobacterial antigens were compared following challenge. IFN-γ responses to mycobacterial antigens were detected at 29 days after challenge and were sustained during the remainder of the study. IFN-γ responses to mycobacterial antigens exceeded corresponding responses in nonstimulated cultures. IP-10 responses to mycobacterial antigens exceeded preinfection responses at 7, 29, and 63 days after challenge. In contrast to IFN-γ responses, IP-10 responses to mycobacterial antigens generally did not exceed the respective responses in nonstimulated cultures. IP-10 responses to medium alone and to mycobacterial antigens followed a similar pattern of response. Correlations between IP-10 and IFN-γ (protein) responses were modest (r≈ 0.50 to 0.65). Taken together, these findings do not support the use of IP-10 protein as a biomarker for bovine tuberculosis using the current testing protocol and reagents; however, mRNA-based assays may be considered for further analysis.


2015 ◽  
Vol 53 (7) ◽  
pp. 2316-2323 ◽  
Author(s):  
Hayley C. King ◽  
Andrew Murphy ◽  
Phillip James ◽  
Emma Travis ◽  
David Porter ◽  
...  

The incidence ofMycobacterium bovis, the causative agent of bovine tuberculosis, in cattle herds in the United Kingdom is increasing, resulting in substantial economic losses. The European badger (Meles meles) is implicated as a wildlife reservoir and is the subject of control measures aimed at reducing the incidence of infection in cattle populations. Understanding the epidemiology ofM. bovisin badger populations is essential for directing control interventions and understanding disease spread; however, accurate diagnosis in live animals is challenging and currently uses invasive methods. Here we present a noninvasive diagnostic procedure and sampling regimen using field sampling of latrines and detection ofM. boviswith quantitative PCR tests, the results of which strongly correlate with the results of immunoassays in the field at the social group level. This method allowsM. bovisinfections in badger populations to be monitored without trapping and provides additional information on the quantities of bacterial DNA shed. Therefore, our approach may provide valuable insights into the epidemiology of bovine tuberculosis in badger populations and inform disease control interventions.


2018 ◽  
Vol 6 (15) ◽  
pp. e00258-18
Author(s):  
Olga Andrievskaia ◽  
Marc-Olivier Duceppe ◽  
Dara Lloyd

ABSTRACT Mycobacterium bovis is the causative agent of bovine tuberculosis, an infectious disease that affects both animals and humans and thus presents a risk to public health and the livestock industry. Here, we report the genome sequences of five Mycobacterium bovis strains that represent major genotype clusters observed in farmed animals and wildlife in Canada.


2017 ◽  
Vol 5 (14) ◽  
Author(s):  
Kerri M. Malone ◽  
Damien Farrell ◽  
Tod P. Stuber ◽  
Olga T. Schubert ◽  
Ruedi Aebersold ◽  
...  

ABSTRACT We report here an update to the reference genome sequence of the bovine tuberculosis bacillus Mycobacterium bovis AF2122/97, generated using an integrative multiomics approach. The update includes 42 new coding sequences (CDSs), 14 modified annotations, 26 single-nucleotide polymorphism (SNP) corrections, and disclosure that the RD900 locus, previously described as absent from the genome, is in fact present.


2017 ◽  
Vol 5 (27) ◽  
Author(s):  
Maxime Branger ◽  
Amandine Hauer ◽  
Lorraine Michelet ◽  
Claudine Karoui ◽  
Thierry Cochard ◽  
...  

ABSTRACT Mycobacterium bovis is the etiologic agent of bovine tuberculosis, a chronic infectious disease affecting livestock, wild animals, and sometimes humans. We report here three draft genome sequences of Mycobacterium bovis strains of spoligotypes SB0821 and SB0134, isolated from wildlife but circulating in wildlife-livestock multihost systems, and SB0121, circulating exclusively in cattle.


2014 ◽  
Vol 21 (5) ◽  
pp. 698-703 ◽  
Author(s):  
Deepanker Tewari ◽  
Ernest Hovingh ◽  
Rick Linscott ◽  
Edmond Martel ◽  
John Lawrence ◽  
...  

ABSTRACTVaccination for Johne's disease with killed inactivated vaccine in cattle herds has shown variable success. The vaccine delays the onset of disease but does not afford complete protection. Johne's disease vaccination has also been reported to interfere with measurements of cell-mediated immune responses for the detection of bovine tuberculosis. Temporal antibody responses and fecal shedding ofMycobacterium aviumsubsp.paratuberculosis, the causative agent of Johne's disease, were measured in 2 dairy cattle herds using Johne's disease vaccine (Mycopar) over a period of 7 years. Vaccination against Johne's disease resulted in positive serumM. aviumsubsp.paratuberculosisantibody responses in both herds, and the responses persisted in vaccinated cattle up to 7 years of age. Some vaccinated animals (29.4% in herd A and 36.2% in herd B) showed no serological reactivity toM. aviumsubsp.paratuberculosis.M. aviumsubsp.paratuberculosis-specific antibody responses were also detected in milk from Johne's disease-vaccinated animals, but fewer animals (39.3% in herd A and 49.4% in herd B) had positive results with milk than with serum samples. With vaccination againstM. aviumsubsp.paratuberculosis, fecal shedding in both dairy herds was reduced significantly (P< 0.001). In addition, when selected Johne's disease-vaccinated and -infected animals were investigated for serological cross-reactivity toMycobacterium bovis, no cross-reactivity was observed.


2018 ◽  
Vol 86 (8) ◽  
Author(s):  
Naoto Noguchi ◽  
Risa Nakamura ◽  
Shinya Hatano ◽  
Hisakata Yamada ◽  
Xun Sun ◽  
...  

ABSTRACT Interleukin 21 (IL-21) is a pleiotropic common cytokine receptor γ chain cytokine that promotes the effector functions of NK cells and CD8+ T cells and inhibits CD8+ T cell exhaustion during chronic infection. We found that the absolute number of short-lived effector CD8+ T cells (SLECs) (KLRG1high CD127low) decreased significantly in IL-21 receptor-deficient (IL-21R−/−) mice during Mycobacterium bovis bacillus Calmette-Guérin (BCG) infection. Early effector CD8+ T cells (EECs) (KLRG1low CD127low) were normally generated in IL-21R−/− mice after infection. Exhausted CD8+ T cells (PD-1high KLRG1low) were also normally generated in IL-21R−/− mice after infection. Mixed bone marrow (BM) chimera and transfer experiments showed that IL-21R on CD8+ T cells was essential for the proliferation of EECs, allowing them to differentiate into SLECs after BCG infection. On the other hand, the number of SLECs increased significantly after infection with recombinant BCG (rBCG) that secreted an antigen 85B (Ag85B)–IL-21 fusion protein (rBCG–Ag85B–IL-21), but the number of exhausted CD8+ T cells did not change after rBCG–Ag85B–IL-21 infection. These results suggest that IL-21 signaling drives the differentiation of SLECs from EECs but does not inhibit the exhaustion of CD8+ T cells following BCG infection in mice.


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