Diagnosis of Mycobacterium bovis infection in livestock using gamma interferon assay and single intradermal comparative tuberculin test in Assam and Meghalaya

Author(s):  
Acheenta G. Barua ◽  
Himangshu Raj ◽  
Ashok Kumar ◽  
Chandana C. Barua ◽  
Arundhati Purkayastha ◽  
...  

The present study was carried out to investigate the diagnostic potential of gamma interferon (IFN-ã) assay and single intradermal comparative tuberculin test (SICTT), including species specification of bovine tuberculosis infection in different livestock farms of Assam and Meghalaya. A total of 199 animals (cattle and buffalo) were examined for bovine tuberculosis symptoms and swab samples were cultured. Biochemical tests and PCR were used for species specification of bovine tuberculosis. Out of 199 cases examined, 33 (16.58%) showed positive for SICTT, 39 (19.59%) for IFN-ã and 35(17.59%) for PCR. Based on PCR targeting pncA region, the confirmation was done for M. Bovis. IFN-ã thus ensures a sensitive and specific detection of early bovine tuberculosis infection together with SICTT and hence may be considered as a screening method of choice.

2006 ◽  
Vol 13 (6) ◽  
pp. 611-619 ◽  
Author(s):  
W. R. Waters ◽  
M. V. Palmer ◽  
T. C. Thacker ◽  
J. B. Payeur ◽  
N. B. Harris ◽  
...  

ABSTRACT Cross-reactive responses elicited by exposure to nontuberculous mycobacteria often confound the interpretation of antemortem tests for Mycobacterium bovis infection of cattle. The use of specific proteins (e.g., ESAT-6, CFP-10, and MPB83), however, generally enhances the specificity of bovine tuberculosis tests. While genes for these proteins are absent from many nontuberculous mycobacteria, they are present in M. kansasii. Instillation of M. kansasii into the tonsillar crypts of calves elicited delayed-type hypersensitivity and in vitro gamma interferon and nitrite concentration responses of leukocytes to M. avium and M. bovis purified protein derivatives (PPDs). While the responses of M. kansasii-inoculated calves to M. avium and M. bovis PPDs were approximately equivalent, the responses of M. bovis-inoculated calves to M. bovis PPD exceeded their respective responses to M. avium PPD. The gamma interferon and nitrite responses of M. kansasii-inoculated calves to recombinant ESAT-6-CFP-10 (rESAT-6-CFP-10) exceeded corresponding responses of noninoculated calves as early as 15 and 30 days after inoculation, respectively, and persisted throughout the study. The gamma interferon and nitrite responses of M. bovis-inoculated calves to rESAT-6-CFP-10 exceeded the corresponding responses of M. kansasii-inoculated calves beginning 30 days after inoculation. By using a lipoarabinomannan-based enzyme-linked immunosorbent assay, specific serum antibodies were detected as early as 50 days after challenge with M. kansasii. By a multiantigen print immunoassay and immunoblotting, serum antibodies to MPB83, but not ESAT-6 or CFP-10, were detected in M. kansasii-inoculated calves; however, responses to MPB83 were notably weaker than those elicited by M. bovis infection. These findings indicate that M. kansasii infection of calves elicits specific responses that may confound the interpretation of bovine tuberculosis tests.


2012 ◽  
Vol 49 (No. 11) ◽  
pp. 406-412 ◽  
Author(s):  
S. I B Cadmus ◽  
N. N Atsanda ◽  
S. O Oni ◽  
E. E U Akang

Bovine tuberculosis was investigated in one private herd with 171 cattle after five cases were suspected to be tuberculous following post mortem examination. Using the intradermal comparative cervical tuberculin test 18 (10.5%) animals (ages from 2 to 12 years) were positive: 11 animals of N’dama breed and seven animals of White Fulani (i.e. Bunaji) breed; 17 female and one male animal. In all 11 randomly selected positive reactors, a spectrum of tuberculous lesions affecting the lungs, spleen, heart, liver, and the lymph nodes were observed. All the smear samples obtained were positive for acid-fast bacilli; cultural isolation confirmed the growth of mycobacteria on pyruvate-enriched Loewenstein-Jensen medium, which were identified by molecular typing to be Mycobacterium bovis. This study demonstrates widespread infection in this cattle herd and potential risk of infection for the human population with M. bovis.


2011 ◽  
Vol 321 (1) ◽  
pp. 30-36 ◽  
Author(s):  
Yang Wang ◽  
Xiangmei Zhou ◽  
Jingjun Lin ◽  
Fei Yin ◽  
Lihua Xu ◽  
...  

2000 ◽  
Vol 68 (10) ◽  
pp. 6073-6076 ◽  
Author(s):  
Timo Ulrichs ◽  
Peter Anding ◽  
Steven Porcelli ◽  
Stefan H. E. Kaufmann ◽  
Martin E. Munk

ABSTRACT Numbers of gamma interferon (IFN-γ)-producing cells reactive to ESAT-6 antigen were increased in recent converters to purified protein derivative positivity and in tuberculosis patients but not in unvaccinated or Mycobacterium bovis BCG-vaccinated healthy donors. ESAT-6-reactive IFN-γ-producing cells in recent converters and tuberculosis patients recognized similar synthetic peptides. Thus, ESAT-6 is a potential candidate for use in detection of early, as well as active, tuberculosis and for control of the disease.


2003 ◽  
Vol 130 (3) ◽  
pp. 485-490 ◽  
Author(s):  
L. COBOS-MARÍN ◽  
J. MONTES-VARGAS ◽  
S. RIVERA-GUTIERREZ ◽  
A. LICEA-NAVARRO ◽  
J. A. GONZÁLEZ-Y-MERCHAND ◽  
...  

Bovine tuberculosis is a zoonotic disease that not only causes huge economic losses but also poses an important risk for human infection. The definitive identification of a clinical isolate relies on time-consuming, highly specialized and laborious biochemical tests. We have developed a method for the rapid and reliable identification of Mycobacterium bovis and for its simultaneous differentiation from other members of the M. tuberculosis complex. Furthermore, the technique also allowed us to distinguish M. tuberculosis complex members from other Mycobacterial species. The method comprises both a single PCR and a multiplex-PCR and can be confidently applied to samples of both veterinary and human origin.


2016 ◽  
Vol 37 (5Supl2) ◽  
pp. 3719
Author(s):  
Antonio Francisco de Souza Filho ◽  
Ana Luiza Alves Rosa Osório ◽  
Klaudia Dos Santos Gonçalves Jorge ◽  
Flábio Ribeiro Araújo ◽  
Carlos Eugênio Soto Vidal ◽  
...  

Mycobacterium bovis is the agent of bovine tuberculosis, a disease endemic to all Brazilian states. Molecular typing techniques help to stratify and refine data, providing information that facilitates epidemiological research. In this study, MIRU-VNTR, targeting 24 loci, was employed to identify and characterize the genetic groups of M. bovis isolates obtained from an outbreak of bovine tuberculosis. Eighteen acid-fast bacilli isolates, obtained from bovine tissue samples, and reactive to the comparative cervical tuberculin test, were identified as species of the M. tuberculosis complex, and were genotyped by MIRU-VNTR with 24 primer pairs. Genotyping revealed three genetic profiles comprising one with 15 isolates (83.3%), one with two isolates (11.1%), and one profile with one unique isolate (5.6%). This distinction was achieved with the MIRU 31 primer, which resulted in clustering of two isolates into the same profile, and ETR A, B, and C, which discriminated the isolate with a unique profile. The occurrence of clustered isolates is indicative of recent transmission, whereas isolates with a unique profile suggest reactivation of latent infection. The presence of different M. bovis genotypes in the same herd suggests movement of infected animals or different sources of intra-herd infection. Use of the MIRU-VNTR molecular epidemiology technique in M. bovis isolates obtained from an outbreak of bovine tuberculosis in Rio Grande do Sul state demonstrated the genetic diversity of circulating strains, despite the presence of a predominant group.


2021 ◽  
Vol 10 (3) ◽  
pp. 229-233

Many diagnostic tools are essential for Mycobacterium bovis (M. bovis) eradication program. This study aimed to apply γ-IFN assay to detect bovine tuberculosis and multiplex PCR (m-PCR) for rapid identification of Mycobacterial isolates. A total no. of 150 cattle in 10 small farms at different Governorates in Egypt, were previously gave suspected results with comparative cervical tuberculin test (SICCT), they retested after 60 days later again with SICCT and bovine gamma-interferon (γ-IFN) immunoassay. Eighty-seven (58%) out of total 150 animals were +ve reactors by SICCT test while 80 (53.3%) animals gave +ve γ-IFN assay. The isolated M. bovis by conventional culturing and identification tests were +ve 55 (63.2%) out 87. The γ-IFN assay sensitivity and specificity gave 82.9% and 93.8% respectively. For rapid identification of different mycobacterial isolates using m-PCR two set of primers were used. The first set gave 123bp DNA PCR product expressing IS6110 insertion element for Mycobacterium tuberculosis (MTBC). The other one gave 500bp from RvD1Rv2031c genomic sequence definite to M. bovis. M-PCR findings were in a concordance with results of conventional culturing and identification tests with high sensitivity and specificity (100%). From this study, it is concluded that diagnosis of bovine tuberculosis (BTB) used tuberculin test and γ-IFN assay with m-PCR for rapid identification M. bovis isolates in living herds.


2016 ◽  
Vol 6 (6) ◽  
pp. 1069
Author(s):  
Paramjeet Ahir ◽  
Gursimran Filia ◽  
Vishal Mahajan ◽  
Geeta D Leishangthem ◽  
Tejinder S Rai ◽  
...  

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.


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