scholarly journals Field Evaluation of a Novel Differential Diagnostic Reagent for Detection of Mycobacterium bovis in Cattle

2006 ◽  
Vol 13 (10) ◽  
pp. 1119-1124 ◽  
Author(s):  
P. J. Cockle ◽  
S. V. Gordon ◽  
R. G. Hewinson ◽  
H. M. Vordermeier

ABSTRACT In the search for improved tools with which to control bovine tuberculosis, the development of enhanced immunodiagnostic reagents is a high priority. Such reagents are required to improve the performance of tuberculin-based reagents and allow the discrimination of vaccinated cattle from those infected with Mycobacterium bovis. In this study, we identified the immunodominant, frequently recognized peptides from Rv3873, Rv3879c, Rv0288, and Rv3019c, which, together with peptides comprising the current lead diagnostic antigens, ESAT-6 and CFP-10, were formulated into a peptide cocktail. In a test of naturally infected cattle, this cocktail was significantly better than tuberculin was for identifying skin test-negative animals with confirmed bovine tuberculosis. In addition, the specificity of this cocktail was not compromised by Mycobacterium bovis BCG vaccination. In summary, our results prioritize this peptide-based, fully synthetic reagent for assessment in larger trials.

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.


2012 ◽  
Vol 19 (5) ◽  
pp. 797-803 ◽  
Author(s):  
S. Flores-Villalva ◽  
F. Suárez-Güemes ◽  
C. Espitia ◽  
A. O. Whelan ◽  
M. Vordermeier ◽  
...  

ABSTRACTThe mycobacterial immunodominant ESAT-6 and CFP-10 antigens are strongly recognizable in tuberculosis-infected cattle, and they do not elicit a response in cattle without infection. In addition, they are absent in most environmental mycobacterial species, and therefore, their use can be an alternative to purified protein derivative (PPD) tuberculin in the development of a more specific skin diagnostic test in cattle. The aim of the current study was to assess the potential of an ESAT-6 and CFP-10 (E6-C10) protein cocktail in a skin test format in naturally tuberculosis-infected and paratuberculosis-infected cattle. We also included MPB83 as a third component in one of the protein cocktail preparations. The protein cocktail was tested at different dose concentrations (5, 10, and 15 μg per protein). The best skin response to the E6-C10 protein cocktail was obtained with 10 μg. Subsequently, this concentration was tested in 2 herds with high and low bovine tuberculosis prevalence, the latter with paratuberculosis coinfection. Our data show that the E6-C10 cocktail allows identification of an important proportion of animals that PPDB is not able to recognize, especially in low-prevalence herds. The protein cocktail did not induce reactions in tuberculosis-free cattle or in paratuberculosis-infected cattle. Addition of MPB83 to the protein cocktail did not make any difference in the skin reaction.


2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Claire Barry ◽  
David Corbett ◽  
Douwe Bakker ◽  
Peter Andersen ◽  
Jim McNair ◽  
...  

Bovine tuberculosis (bTB) is diagnosed in naturally infected populations exposed to a wide variety of other pathogens. This study describes the cell-mediated immune responses of cattle exposed toMycobacterium aviumsubspeciesparatuberculosis(Map) andMycobacterium aviumsubspeciesaviumwith particular reference to routine antefmortemMycobacterium bovisdiagnostic tests. The IFN-γreleased in response to stimulated blood was found to peak later in the Map-exposed group and was more sustained when compared to the Maa-exposed group. There was a very close correlation between the responses to the purified protein derivatives (PPD) used for stimulation (PPDa, PPDb, and PPDj) with PPDa and PPDj most closely correlated. On occasion, in the Map-infected cattle, PPDb-biased responses were seen compared to PPDa suggesting that some Map-infected cattle could be misclassified asM. bovisinfected using this test with these reagents. This bias was not seen when PPDj was used. SICCT results were consistent with the respective infections and all calves would have been classed skin test negative.


2007 ◽  
Vol 15 (1) ◽  
pp. 168-171 ◽  
Author(s):  
Jose Domínguez ◽  
Juan Ruiz-Manzano ◽  
Malú De Souza-Galvão ◽  
Irene Latorre ◽  
Celia Milà ◽  
...  

ABSTRACT We evaluated the T-SPOT.TB and Quantiferon-TB Gold In tube (QFN-G-IT) tests for diagnosing Mycobacterium tuberculosis infection. T-SPOT.TB was more sensitive than QFN-G-IT in diagnosing both active and latent infection. Both gamma interferon tests were unaffected by prior Mycobacterium bovis BCG vaccination. Among children who were not BCG vaccinated but had a positive tuberculin skin test, QFN-G-IT was negative in 53.3% of cases, and T-SPOT.TB was negative in 50% of cases.


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.


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.


2018 ◽  
Vol 30 (3) ◽  
pp. 468-470 ◽  
Author(s):  
Salwa Al-Mouqatea ◽  
Mohammad Alkhamis ◽  
Batool Akbar ◽  
Abdulmohsen Ali ◽  
Hameed Al-Aqeel ◽  
...  

Bovine tuberculosis (TB) is endemic in Kuwait; cattle identified as TB-positive using the caudal fold test (CFT) are culled. We used a Bayesian approach to estimate the sensitivity (Se) and specificity (Sp) of the IFNγ assay and ELISA, which are not routinely used in Kuwait in CFT-negative dairy cattle. Blood samples from CFT-negative cattle ( n = 384) collected from 38 dairy farms were tested by IFNγ assay and ELISA. The Se and Sp (95% CI) of the IFNγ were 85.0% (67.6–95.3%) and 90.4% (86.7–95.3%), respectively, whereas estimates for the ELISA were 61.1% (33.1–84.6%) and 85.4% (81.7–88.8%). TB prevalence (95 CI%) in CFT-negative cattle was estimated as 2.6% (0.5–9.5%). The IFNγ assay may play a role as an ancillary test for the identification of Mycobacterium bovis–infected cattle that are undetected by CFT.


2013 ◽  
Vol 33 (12) ◽  
pp. 1403-1408
Author(s):  
Mayara Fernanda Maggioli ◽  
Joyce Rodrigues Lobo ◽  
Maria Clorinda Soares Fioravanti ◽  
André Kipnis ◽  
Ana Paula Junqueira-Kipnis

The present study aimed to assess the CD4, CD8 and γδ blood levels for Curraleiro Pé-duro, as well as the specific IFN-γ response after BCG vaccination using flow cytometry. The specific immune response against BCG was also evaluated by tuberculin skin test, performed before and 45 days after the vaccination. For comparison purposes, the same parameters were investigated on Nellore calves, an exotic bovine with resistance previously demonstrated. Naturally, Curraleiro Pé-duro animals had greater levels of CD4, CD8 and γδ lymphocytes (p<0.05). In response to vaccine, Curraleiro Pé-duro showed greater ability to respond specifically to BCG, generating resistance profile (Th1), evidenced by greater number of antigen specific CD4+ cells producing IFN-γ (p<0.05) and also higher tuberculin skin test reaction (p<0.05). Additionally, vaccinated Curraleiro Pé-duro calves had higher CD4 cells numbers than both Nellore control (p<0.05) and vaccinated groups (p<0.05). Curraleiro Pé-duro calves' higher basal lymphocytes blood level and stronger response in both IFN-γ and tuberculin skin test parameters probably play a positive role on protection/resistance to Mycobacterium bovis.


2017 ◽  
Author(s):  
L. McCallan ◽  
C. Brooks ◽  
C. Couzens ◽  
F. Young ◽  
A.W. Byrne ◽  
...  

AbstractThe ability to accurately identify infected hosts is the cornerstone of effective disease control and eradication programs. In the case of bovine tuberculosis, caused by infection with the pathogenMycobacterium bovis, accurately identifying infected individual animals has been challenging as all available tests exhibit less than 100% discriminatory ability. Here we assess the utility of three serological tests and assess their performance relative to skin test (Single Intradermal Comparative Cervical Tuberculin; SICCT), gamma-interferon (IFNγ) and post-mortem results in a Northern Ireland setting. Furthermore, we describe a case-study where one test was used in conjunction with statutory testing.Serological tests using samples taken prior to SICCT disclosed low proportions of animals as test positive (mean 3% positive), despite the cohort having high proportions with positive SICCT test under standard interpretation (121/921; 13%) or IFNγ (365/922; 40%) results. Furthermore, for animals with a post-mortem record (n=286), there was a high proportion with TB visible lesions (27%) or with laboratory confirmed infection (25%). As a result, apparent sensitivities within this cohort was very low (≤15%), however the tests succeeded in achieving very high specificities (96-100%). During the case-study, 7/670 (1.04%) samples from SICCT negative animals from a large chronically infected herd were serology positive, with a further 10 animals being borderline positive (17/670; 2.54%). 9/17 of these animals were voluntarily removed, none of which were found to be infected (-lesions/-bacteriology) post-mortem; 1 serology test negative animal was subsequently lesion+ andM bovisconfirmed at slaughter.ImportanceEradication of bovine tuberculosis (bTB; caused byMycobacterium bovis) has remained elusive in a number of countries despite long-term coordinated test and cull programs. This can partially be explained by the limitations of available statutory tests; therefore supplementary test platforms that identify additional infected animals would be of significant utility. Overall, during our study three serological tests did not disclose a high proportion of animals as infected in high-risk cattle herds, and exhibited limited ability to disclose animals that were positive to the statutory skin test, the gamma interferon test (IFNγ), or were post-mortem confirmed withM. bovis. These serological tests could be used in a supplementary fashion to the statutory tests in particular circumstances; but may be of limited advantage where parallel use of IFNγ and skin testing is performed, as these tests together tended to disclose the majority of animals with post-mortem evidence of infection in our study cohort.


2016 ◽  
Vol 37 (2) ◽  
pp. 885
Author(s):  
Silvia Minharro ◽  
Cristiane De Morais Alves ◽  
Pedro Moacyr Pinto Coelho Mota ◽  
Elaine Maria Seles Dorneles ◽  
Andréa Padilha de Alencar ◽  
...  

Water buffalo are of great economic importance in Brazilian Amazonia, which has the largest herd in Brazil. Bovine tuberculosis caused by Mycobacterium bovis is a zoonotic disease that results in severe losses to water buffalo production. Although the disease has already been described in the country, data on the occurrence and distribution of bovine tuberculosis in water buffalo in Brazil is very scarce. The aim of this study was to determine the frequency of bovine tuberculosis in water buffalo in the Baixo Araguari River region of Amapá, Brazil. Thirty herds, randomly selected from the 41 herds of water buffalo in the region, were sampled. From those herds, 212 randomly selected water buffalo were subjected to the comparative tuberculin skin test. The proportion of Baixo Araguari River region herds that were positive for bovine tuberculosis was 50.0% (95% CI 31.3% to 68.7%) and the proportion of animals that were positive was estimated to be 14.8% (95% CI 7.8% to 21.9%). Our results show that bovine tuberculosis is spread widely among water buffalo in the Baixo Araguari River region, which suggests that measures to control the disease should be undertaken in the region.


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