Gamma-interferon assay for the ancillary diagnosis of bovine tuberculosis in dairy cattle in urban and adjacent areas of Dhaka city, Bangladesh.

Background and Aim: Bovine tuberculosis (bTB) is an infectious disease of cattle, mainly caused by Mycobacterium bovis. This study aimed to evaluate the efficacy of the interferon-gamma (IFN-γ) assay and single-intradermal comparative tuberculin test (SICTT) in detecting bTB. Materials and Methods: In an earlier study, 150 positive, 83 inconclusive, and 480 negative animals from 24 cattle herds were screened using SICTT. From these groups, 125 positive, 17 inconclusive, and six negative animals were subsequently verified using the IFN-γ assay. Single-intradermal comparative tuberculin test outcomes were interpreted according to standard guidelines, whereas blood samples were collected and stimulated with purified protein derivatives. Sandwich enzyme-linked immunosorbent assay was used to measure secreted IFN-γ. Concordant and Bayesian latent class analyses were performed to evaluate test performance. Results: Results from the IFN-γ assay revealed that 83.2%, 64.7%, and 16.67% of the animals were positive in the SICTT-positive, inconclusive, and negative animal categories, respectively. Sensitivity (SE) and specificity (SP) of SICTT were 83.9% (95% confidence interval [CI]: 77.4-90.1) and 95.7% (95% CI: 86.9-99.7), respectively. Sensitivity and SP for the IFN-γ assay were 78.9% (95% CI: 71.9-85.4) and 83.9% (65.9-95.9), respectively. The use of both tests in parallel increases the SE of bTB detection (~94%), compared with SICTT alone. Conclusion: Use of the IFN-γ assay with SICTT in parallel, predominantly on cattle demonstrating an inconclusive SICTT outcome, boosts bTB detection rate in low resource settings.

Enhancing bovine tuberculosis screening at Dhaka city in Bangladesh: Integrating gamma interferon blood test as ancillary testing with tuberculin skin test.

Tuberculin skin test (TST) is the standard method for screening of bovine tuberculosis (bTB). However, gamma interferon blood test has been introduced in the bTB control program as an ancillary testing with TST in many countries of the world. The objective of this study was to recommend this screening test as an ancillary testing with TST for field application in Bangladesh. In this study 577 cattle of different age, sex and breeds from twenty nine (29) cattle herds were examined to determine skin response against bTB through single intradermal comparative tuberculin test (SICTT) that comprised of positive (n = 81), inconclusive (n = 44) and negative (n = 452) animals. Of which 74 animals that included positive (n = 63), inconclusive (n = 8) and negative (n = 3) animals were taken under this study. Blood samples were collected in heparinized tube and stimulated overnight with bovine and avian purified protein derivatives (PPDs) for the secretion of gamma interferon, and measured via sandwich ELISA. Cohen's kappa statistics was performed for the evaluation of agreement between the two tests. The agreement obtained between two tests was fair (Kappa agreement, K = 24.0%, 95% CI = 16.9-30.5%, P = 0.037). Of positive (n = 63), inconclusive (n = 8) and negative (n = 3) status of animals at SICTT, 82.54% (n = 52), 62.50% (n = 5), and 33.33% (n = 1) were found to be bTB positive respectively through this ancillary test. This test notably corroborates to TST result. A considerable number of inconclusive TB status animals were found to be positive through this gamma interferon assay. Therefore, this test could be used as an ancillary test with TST to maximize the proportion of bTB estimation in the infected cattle herd for early detection of zoonotic tuberculosis in Bangladesh before transmission at the animal-human interface.

Optimisation of the tuberculin skin test for detection of Mycobacterium bovis in African buffaloes (Syncerus caffer).

Effective screening methods are critical for preventing the spread of bovine tuberculosis (bTB) among livestock and wildlife species. The tuberculin skin test (TST) remains the primary test for bTB globally, although performance is suboptimal. African buffaloes (Syncerus caffer) are a maintenance host of Mycobacterium bovis in South Africa, tested using the single intradermal tuberculin test (SITT) or comparative test (SICTT). The interpretation of these tests has been based on cattle thresholds due to the lack of species-specific cut-off values for African buffaloes. Therefore, the aims of this study were to calculate buffalo-specific thresholds for different TST criteria (SITT, SICTT, and SICTT72h that calculates the differential change at 72 h only) and compare performance using these cut-off values. The results confirm that 3 mm best discriminates M. bovis-infected from unexposed control buffaloes with sensitivities of 69 % (95 % CI 60-78; SITT and SICTT) and 76 % (95 % CI 65-83; SICTT72h), and specificities of 86 % (95 % CI 80-90; SITT), 96 % (95 % CI 92-98; SICTT72h) and 97 % (95 % CI 93-99; SICTT), respectively. A comparison between TST criteria using buffalo-specific thresholds demonstrates that the comparative TST performs better than the SITT, although sensitivity remains suboptimal. Therefore, further research and the addition of ancillary tests, such as cytokine release assays, are necessary to improve M. bovis detection in African buffaloes.

Characteristics of Mycobacterium bovis infected herds tested with the interferon-gamma assay.

The IFN-γ (interferon gamma) assay is used in Ireland as an ancillary diagnostic test to the single intradermal comparative tuberculin test (SICTT) to maximise the detection of Mycobacterium bovis infected animals (bTB) in cattle herds. Understanding the relationships between herd and animal risk factors and IFN-γ test results is critical to enable the development and evaluation of policy measures on how best to use the test. In this study, we set out to characterise Irish herds with IFN-γ test positive animals in terms of herd size, number of SICTT reactors and number of IFN-γ positive tests, and to evaluate the IFN-γ test in terms of the test cut-off values. The results showed that larger herds with more SICTT reactors were likely to have more IFN-γ positives in the herd, and herds with an IFN-γ test positive animal that was also positive for bTB lesions at post-mortem had higher numbers of IFN-γ positive animals in the herd. Raising the cut-off values for the IFN-γ test only marginally decreased the combined sensitivity of the IFN-γ and the SICTT for diagnosis of bTB lesioned animals. The analysis has provided valuable information on the performance of the IFN-γ test as it is used under current bTB infection levels in Ireland.

Bayesian latent class estimation of sensitivity and specificity parameters of diagnostic tests for bovine tuberculosis in chronically infected herds in Northern Ireland.

In the European Union, the recommended ante-mortem diagnostic methods for bovine tuberculosis (bTB) include the single intradermal cervical comparative tuberculin (SICCT) test and the interferon-gamma (IFN-γ) test as an ancillary test. The SICCT test has a moderate sensitivity (Se) and high specificity (Sp), while the IFN-γ test has good Se, but a lower Sp than the SICCT test. A retrospective Bayesian latent class analysis was conducted on 71,185 cattle from 806 herds chronically infected with bTB distributed across Northern Ireland (NI) to estimate the Se and Sp of the common ante-mortem tests and meat inspection. Analyses were also performed on data stratified by farming type and herd location to explore possible differences in test performance given the heterogeneity in the population. The mean estimates in chronically infected herds were: (1) 'standard' SICCT: Se 40.5-57.7%, Sp 96.3-99.7%; (2) 'severe' SICCT: Se 49.0%-60.6%, Sp 94.4-99.4%; (3) IFN-γ(bovine-avian) using a NI optical density (OD) cut-off difference of 0.05: IFN-γ(B-A)NI: Se 85.8-93.0%, Sp 75.6-96.2%; (4) IFN-γ(bovine-avian) using a standard 'commercial' OD cut-off difference of 0.1: IFN-γ(B-A)0.1: Se 83.1-92.1%, Sp 83.1-97.3%; and (5) meat inspection: Se 49.0-57.1% Se, Sp 99.1-100%. Se estimates were lower in cattle from dairy farms than from beef farms. There were no notable differences in estimates by location of herds. Certain population characteristics, such as production type, might influence the ability of bTB tests to disclose truly infected cases.

Prevalence of bovine tuberculosis in cattle, goats, and camels of traditional livestock raising communities in Eritrea.

BACKGROUND: The aim of the current study was to assess the prevalence of bovine tuberculosis (BTB) in cattle, goats, and camels, and its zoonotic potential within the traditional livestock raising communities in four regions of Eritrea. The Single Intradermal Comparative Tuberculin Test (SICTT) as indicator of M. bovis infection was conducted on 1077 cattle, 876 goats, and 195 camels. To elucidate possible risk factors for BTB transmission between animals and its potential zoonotic implication, questionnaire based face-to-face interviews were conducted in households of which 232 raised cattle, 128 goats, and 29 camels. RESULTS: The results of the SCITT were interpreted using the OIE standard (> 4 mm cut-off) for positive responses. In cattle, individual animal (n = 1077) and herd (n = 413) prevalences were 1.2% (n = 13) [Confidence Interval (CI) 95% CI, 1.0-1.3%] and 3.2% (n = 13) (95% CI, 3.0-3.4%), respectively. In goats (n = 876), none of the animals was positive. In camels, individual animal (n = 195) and herd (n = 70), BTB prevalences were 1.5% (n = 3) (95% CI,1.4-1.6%) and 2.9(n = 2) (95% CI, 0.9-4.6%), respectively. Overall, male animals were more at risk (OR = 2.6; 95% CI:1.0-8.7) when compared to females. Sharing of water points, introduction of new animals into herds and migration of animals over large distances were common events that may contribute to intra and inter-species transmission of BTB. Consumption of raw milk, lack of BTB transmission awareness, and low levels of education were common in the farming communities. CONCLUSION: The current study highlighted a low prevalence of M. bovis in cattle, goats and camels in extensive traditional livestock in Eritrea. Despite this, the spatial distribution of affected animals across most of the sampled regions and consumption of unpasteurized milk warrants surveillance, cautious and timely control measures for the disease.

The performance of the interferon gamma assay when used as a diagnostic or quality assurance test in Mycobacterium bovis infected herds.

There are two different contexts in the Irish bTB eradication programme in which the interferon-gamma assay (IFN-γ) is applied. Firstly, the IFN-γ assay is applied routinely to high risk cohorts in herds with four or more reactors to the SICTT. The IFN-γ test is then carried out on blood samples submitted to the laboratory within 8h of collection (diagnostic testing). Secondly, the use of the IFN-γ assay has recently been extended to test SICTT reactors as part of a general quality assurance (QA) scheme to monitor the performance of the SICTT. Blood samples from reactors are tested one day after blood collection (QA testing). In this study, we analysed the relative performance of the SICTT and IFN-γ when used in parallel as an 8h diagnostic test and as a 24h QA test on SICTT reactors. A total of 17,725 IFN-γ tests were included in the analysis (11,658 diagnostic tests and 6067 QA tests). Of the samples submitted for diagnostic testing, the proportion positive to IFN-γ decreased with the severity of interpretation of the SICTT result. Of the standard reactors that were tested with IFN-γ in the QA programme, 92.2% were positive to the IFN-γ test. Among animals that were SICTT -ve/IFN-γ +ve, 18.9% were positive at post-mortem compared to 11.8% of those that were SICTT +ve (standard reactor)/IFN-γ -ve. These results highlight the risk associated with retaining SICTT -ve/IFN-γ +ve animals, and suggest that prompt removal of these animals is necessary to reduce the potential for future transmission.

Prevalence and risk factors of bovine tuberculosis in dairy cattle in Eritrea.

BACKGROUND: The prevalence of bovine tuberculosis (BTB) in dairy cattle in the three major milk producing regions of Eritrea was assessed by subjecting 15,354 dairy cattle, 50 % of Eritrea's dairy cattle population, to the single intradermal comparative tuberculin test (SICTT). Skin test results were interpreted according to guidelines of the World Organization for Animal Health (OIE) with >4 mm as cutoff in skin thickness increase. In addition, we studied the relation between 'physiological' variables related to pregnancy and lactation, and the variable 'region' on the probability to be skin test positive. RESULTS: The BTB prevalences at animal and herd levels were: 21.5% and 40.9% in Maekel, 7.3% and 10% in Debub, and 0.2% and 1.6% in the Anseba region, respectively. Overall, in the regions included, prevalence was 11.3% (confidence interval (CI) 95% CI, 11.29 - 11.31%) and 17.3% (95% CI, 17.27-17.33%), at animal and herd level, respectively. Considering positive herds only, the animal BTB prevalence was 36.8%, 30.1%, and 1.8%, in Maekel, Debub and Anseba, respectively, and the overall animal prevalence within these herds was 32%. In adult dairy cattle the probability of positive reactivity in the SICTT test was highest in pregnant animals as compared to the other categories. CONCLUSION: This study reports persistent prevalence of BTB as defined by positive SICTT in the dairy sector of Eritrea, especially in the regions of Maekel and Debub that are located in the central highlands of the country. To our understanding this is the first report that has encompassed all the major dairy farms in Eritrea and it will be instrumental in advocating future BTB control programs in the dairy sector.

Parameter estimation and use of gamma interferon assay for the diagnosis of bovine tuberculosis in Brazil / Estimação dos parâmetros e uso do teste do interferon gama para o diagnóstico da tuberculose bovina no Brasil

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.(AU)

O presente estudo avalia a interferência do teste de tuberculinização no teste do interferon gama (INFg), estima a sensibilidade e a especificidade do INFg em condições brasileiras e simula a utilização dos testes múltiplos usando a tuberculinização comparada e o teste do INFg. Trezentos e cinquenta animais oriundos de dois rebanhos livres e dois rebanhos positivos foram submetidos à tuberculinização comparada e ao teste de INFg. A tuberculinização comparada foi realizada utilizando PPD aviária e bovina. O teste de INFg foi realizado utilizando o kit Bovigam® (CSL Veterinary, Austrália) de acordo com as especificações do fabricante. A sensibilidade e especificidade do teste de INFg foram calculadas pelo Modelo Bayesiano de Classe Latente. Esses parâmetros foram também estimados para os testes múltiplos. Os resultados do teste de INFg no D0 e D3 após o teste de tuberculinização foram comparados pelos testes estatísticos de McNemar e kappa. Os resultados das médias de densidade ótica do teste de INFg em ambos os dias foram similares. A sensibilidade e a especificidade do teste de INFg apresentaram resultados variando (95% intervalo de confiança) de 72 a 100% e 74 a 100%, respectivamente. A sensibilidade do teste em paralelo foi acima de 97,5% enquanto a especificidade do teste em série foi acima de 99,7%. O teste do INFg provou ser um método de diagnóstico muito útil.(AU)