scholarly journals Using of Gamma Interferon γ-IFN and Multiplex PCR (m-PCR) for Detection of Bovine Tuberculosis in Dairy Herds in Egypt

2021 ◽  
Vol 10 (3) ◽  
pp. 229-233
Keyword(s):  
Multiplex Pcr ◽  
Sensitivity And Specificity ◽  
Bovine Tuberculosis ◽  
Genomic Sequence ◽  
Tuberculin Test ◽  
Interferon Γ ◽  
Rapid Identification ◽  
Gamma Interferon ◽  
Diagnostic Tools ◽  
Assay Sensitivity

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.

scholarly journals Evaluation of Mycobacterium tuberculosis lipoarabinomannan antigen assay and rapid serology blood test for the diagnosis of bovine tuberculosis in Ethiopia

2019 ◽  
Vol 15 (1) ◽  
Author(s):  
Aboma Zewude ◽  
Temesgen Mohammed ◽  
Lemma Terfassa ◽  
W. Garrett Hunt ◽  
Xueliang Pan ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Gold Standard ◽  
Bovine Tuberculosis ◽  
Blood Test ◽  
Point Of Care ◽  
Rapid Test ◽  
Tuberculin Test ◽  
Receiver Operating Curve ◽  
Diagnostic Tools ◽  
Ifn Γ

Abstract Background Bovine tuberculosis (bTB) is prevalent in dairy cattle in Ethiopia. Currently used diagnostic tools such as the single intradermal comparative tuberculin test (SICTT) are time consuming and labor intensive. A rapid, easy-to-use and cost-effective diagnostic test would greatly contribute to the control of bTB in developing countries like Ethiopia. In the present study, two point-of-care diagnostic tests were evaluated for the detection of bTB: LIONEX® Animal TB Rapid test, a membrane-based test for the detection of antibodies to Mycobacterium bovis in blood and ALERE® Determine TB Lipoarabinomannan (LAM) Ag, an immunoassay for the detection of lipoarabinomannan (LAM) antigen (Ag) of mycobacteria in urine. A combination of the SICTT and gamma interferon (IFN-γ) test was used as the gold standard for the validation of these point-of-care tests, as it was not feasible to slaughter the study animals to carry out the historical gold standard of mycobacterial culture. A total of 175 heads of cattle having three different bTB infection categories (positive SICTT, negative SICTT, and unknown SICTT status) were used for this study. Result The sensitivity and specificity of TB LAM Ag were 72.2% (95% CI = 62.2, 80.4) and 98.8% (95% CI = 93.6, 99.7), respectively, while the sensitivity and specificity of the LIONEX Animal TB rapid test assay were 54% (95% CI = 44.1 64.3) and 98.8% (95% CI = 93.6, 99.7) respectively. The agreement between TB LAM Ag and SICTT was higher (κ = 0.85; 95% CI = 0.65–0.94) than between TB LAM Ag and IFN-γ (κ = 0.67; 95% CI = 0.52–0.81). The agreement between LIONEX Animals TB Rapid blood test and SICTT was substantial, (κ = 0.63; 95% CI = 0.49–0.77) while the agreement between LIONEX Animal TB rapid blood test and IFN-γ test was moderate (κ = 0.53; 95% CI = 0.40–0.67). Analysis of receiver operating curve (ROC) indicated that the area under the ROC curve (AUC) for TB LAM Ag was 0.85 (95% CI = 0.79–0.91) while it was 0.76 (95% CI; =0.69–0.83) for LIONEX Animal TB rapid test assay. Conclusion This study showed that TB LAM Ag had a better diagnostic performance and could potentially be used as ancillary either to SICTT or IFN-γ test for diagnosis of bTB.

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

2017 ◽  
Author(s):  
Acheenta G. Barua ◽  
Himangshu Raj ◽  
Ashok Kumar ◽  
Chandana C. Barua ◽  
Arundhati Purkayastha ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Bovine Tuberculosis ◽  
Screening Method ◽  
Tuberculin Test ◽  
Gamma Interferon ◽  
Tuberculosis Infection ◽  
Specific Detection ◽  
Biochemical Tests ◽  
Diagnostic Potential ◽  
Pcr Targeting

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.

scholarly journals A novel multiplex-PCR for the rapid identification of Mycobacterium bovis in clinical isolates of both veterinary and human origin

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 ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Clinical Isolate ◽  
Mycobacterium Bovis ◽  
Bovine Tuberculosis ◽  
Rapid Identification ◽  
Human Infection ◽  
Economic Losses ◽  
Human Origin ◽  
Biochemical Tests ◽  
Reliable Identification

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.

scholarly journals Nasal swab real-time PCR is not suitable for in vivo diagnosis of bovine tuberculosis

2017 ◽  
Vol 37 (6) ◽  
pp. 549-554
Author(s):  
Fabiana Q. Mayer ◽  
Emily M. dos Reis ◽  
André Vinícius A. Bezerra ◽  
Rogério O. Rodrigues ◽  
Thais Michel ◽  
...  
Keyword(s):  
Real Time ◽  
Sensitivity And Specificity ◽  
Real Time Pcr ◽  
Bovine Tuberculosis ◽  
Tuberculin Test ◽  
Disease Diagnosis ◽  
Economic Losses ◽  
Nasal Swabs ◽  
In Vivo Diagnosis

ABSTRACT: Bovine tuberculosis (bTB) is a zoonosis causing economic losses and public health risks in many countries. The disease diagnosis in live animals is performed by intradermal tuberculin test, which is based on delayed hypersensitivity reactions. As tuberculosis has complex immune response, this test has limitations in sensitivity and specificity. This study sought to test an alternative approach for in vivo diagnosis of bovine tuberculosis, based on real-time polymerase chain reaction (PCR). DNA samples, extracted from nasal swabs of live cows, were used for SYBR® Green real-time PCR, which is able to differentiate between Mycobacterium tuberculosis and Mycobacterium avium complexes. Statistical analysis was performed to compare the results of tuberculin test, the in vivo gold standard bTB diagnosis method, with real-time PCR, thereby determining the specificity and sensitivity of molecular method. Cervical comparative test (CCT) was performed in 238 animals, of which 193 had suitable DNA from nasal swabs for molecular analysis, as indicated by amplification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene, and were included in the study. In total, 25 (10.5%) of the animals were CCT reactive, of which none was positive in the molecular test. Of the 168 CCT negative animals, four were positive for M. tuberculosis complex at real time PCR from nasal swabs. The comparison of these results generated values of sensitivity and specificity of 0% and 97.6%, respectively; moreover, low coefficients of agreement and correlation (-0.029 and -0.049, respectively) between the results obtained with both tests were also observed. This study showed that real-time PCR from nasal swabs is not suitable for in vivo diagnosis of bovine tuberculosis; thus tuberculin skin test is still the best option for this purpose.

scholarly journals Parameter estimation and use of gamma interferon assay for the diagnosis of bovine tuberculosis in Brazil

2012 ◽  
Vol 32 (4) ◽  
pp. 279-283 ◽  
Author(s):  
Luciano B. Lopes ◽  
Telma M. Alves ◽  
Ana Paula R. Stynen ◽  
Pedro M.P.C. Mota ◽  
Rômulo C. Leite ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Multiple Testing ◽  
Latent Class ◽  
Latent Class Model ◽  
Tuberculin Test ◽  
Gamma Interferon ◽  
Kappa Statistics ◽  
Class Model ◽  
Parallel Testing ◽  
Serial Testing

This study aimed to evaluate the interference of tuberculin test on the gamma-interferon (INFg) assay, to estimate the sensitivity and specificity of the INFg assay in Brazilian conditions, and to simulate multiple testing using the comparative tuberculin test and the INFg assay. Three hundred-fifty cattle from two TB-free and two TB-infected herds were submitted to the comparative tuberculin test and the INFg assay. The comparative tuberculin test was performed using avian and bovine PPD. The INFg assay was performed by the BovigamTM kit (CSL Veterinary, Australia), according to the manufacturer's specifications. Sensitivity and specificity of the INFg assay were assessed by a Bayesian latent class model. These diagnostic parameters were also estimate for multiple testing. The results of INFg assay on D0 and D3 after the comparative tuberculin test were compared by the McNemar's test and kappa statistics. Results of mean optical density from INFg assay on both days were similar. Sensitivity and specificity of the INFg assay showed results varying (95% confidence intervals) from 72 to 100% and 74 to 100% respectively. Sensitivity of parallel testing was over 97.5%, while specificity of serial testing was over 99.7%. The INFg assay proved to be a very useful diagnostic method.

Diagnosis of Bovine Tuberculosis in Lactating Cattle and Buffaloes by Comparative Intradermal Tuberculin Test and Bovine Gamma-Interferon Immunoassay

2016 ◽  
Vol 6 (6) ◽  
pp. 1069
Author(s):  
Paramjeet Ahir ◽  
Gursimran Filia ◽  
Vishal Mahajan ◽  
Geeta D Leishangthem ◽  
Tejinder S Rai ◽  
...  
Keyword(s):  
Bovine Tuberculosis ◽  
Tuberculin Test ◽  
Gamma Interferon

Field evaluation of the single intradermal cervical tuberculin test and the interferon-γ assay for detection and eradication of bovine tuberculosis in Spain

1999 ◽  
Vol 70 (1-2) ◽  
pp. 55-66 ◽  
Author(s):  
O.R.González Llamazares ◽  
C.B.Gutiérrez Martı́n ◽  
D.Alvarez Nistal ◽  
V.A.de la Puente Redondo ◽  
L.Domı́nguez Rodrı́guez ◽  
...  
Keyword(s):  
Bovine Tuberculosis ◽  
Tuberculin Test ◽  
Field Evaluation ◽  
Interferon Γ

scholarly journals Bovine tuberculosis: diagnosis in dairy cattle through the association of analyzes

2020 ◽  
Vol 40 (1) ◽  
pp. 12-16
Author(s):  
Leonardo Luiz Dametto ◽  
Ezequiel Davi dos Santos ◽  
Luciana R. Santos ◽  
Elci L. Dickel
Keyword(s):  
Dairy Cattle ◽  
Bovine Tuberculosis ◽  
Clinical Signs ◽  
Tuberculin Test ◽  
Lung Parenchyma ◽  
Diagnostic Tools ◽  
Histopathological Analysis ◽  
Caseous Necrosis ◽  
Central Calcification ◽  
And Control

ABSTRACT: Tuberculosis is a chronic anthropozoonosis of worldwide occurrence, caused by the bacterium Mycobacterium tuberculosis and its variants. In Brazil, the National Program for the Control and Eradication of Brucellosis and Tuberculosis in cattle, is responsible for diagnosing and the correctly allocate positive animals, but there is still a lack of definitive diagnosis of the disease. This study described the use of five diagnostic tools that can be used, preferably together, for the confirmation of suspected cases. These tools included the clinical examination comparative cervical tuberculin test, macroscopic findings during the slaughtering and histopathology of the damaged tissues followed by histochemistry. We evaluated a total of 211 dairy cattle, where 15.1% (32/211) had classic clinical signs of bovine tuberculosis, 74 (35%) showed reactivity in the comparative cervical tuberculin test. Of the total number of animals, 141 (66.8%) were referred for sanitary slaughter due to legal and control issues in the outbreaks of the disease. In the follow-up of slaughtering and inspection of viscera and carcasses, 74 (52.5%) had macroscopic lesions compatible with bovine tuberculosis, while 67 (47.5%) showed no visible changes. During the inspection, fragments of lymph nodes and liver and lung parenchyma were collected from five cattle with macroscopic lesions and five with no lesions. The histopathological analysis showed numerous areas of caseous necrosis with or without central calcification and granulomatous inflammatory infiltrate. In the special staining of Ziehl-Neelsen, numerous acid-fast bacilli were evidenced in all cases.

Assessment of the sensitivity of the gamma-interferon test and the single intradermal comparative cervical test for the diagnosis of bovine tuberculosis under field conditions

2014 ◽  
Vol 143 (1) ◽  
pp. 157-166 ◽  
Author(s):  
A. PRAUD ◽  
M. L. BOSCHIROLI ◽  
L. MEYER ◽  
B. GARIN-BASTUJI ◽  
B. DUFOUR
Keyword(s):  
Bovine Tuberculosis ◽  
Tuberculin Test ◽  
Gamma Interferon ◽  
Skin Tests ◽  
Economic Losses ◽  
The European Union ◽  
Testing Scheme ◽  
Serial Testing ◽  
Significant Difference ◽  
The Individual

SUMMARYIn some French départements, the eradication of bovine tuberculosis is incomplete and usual skin tests [single intradermal tuberculin test (SIT) and single intradermal comparative cervical test (SICCT)] have poor specificity due to cross-reactions with non-pathogenic mycobacteria, causing economic losses. In Côte d'Or (Burgundy, France), an experimental serial testing scheme based on the combination of SICCT and gamma-interferon (IFN-γ) tests has been initiated in order to shorten the interval between suspicion and its invalidation in herds with false-positive results to skin tests. Our aim was to assess the scheme's sensitivity and to compare it to the sensitivity of the screening scheme recommended by the European Commission. Our study included 1768 animals from Côte d'Or. The sensitivities of both schemes were estimated using a Bayesian approach. The individual sensitivity of the IFN-γtest [88·1%, 95% credibility interval (CrI) 72·8–97·5] was not significantly different from individual SICCT sensitivity (80·3%, 95% CrI 61·6–98·0) and individual SIT sensitivity (84·2%, 95% CrI 59·0–98·2). The individual specificity of the IFN-γtest was 62·3% (95% CrI 60·2–64·5). No significant difference could be demonstrated between the sensitivities of the serial testing scheme used in Côte d'Or (73·1%, 95% CrI 41·1–100) and the European Union serial testing scheme (70·1%, 95% CrI 31·5–100·0).

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