Detection of Mycobacterium bovis infection in cattle using an immunoassay for bovine soluble interleukin-2 receptor-α (sIL-2R-α) produced by peripheral blood T-lymphocytes following incubation with tuberculin PPD

O'Nualláin, E M; Davis, William C.; Costello, Eamon; Pollock, J.M.; Monaghan, M. L.

doi:10.1016/s0165-2427(96)05734-0

Cited by 13 publications

(4 citation statements)

References 20 publications

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“…In this study, we confirm the previously observed [13], [16] important roles of IFN- (IFNG), TNF- (TNF), and iNOS (NOS2A) in the bTB immune response. As seen in Figure 2, IL2RA gene expression is significantly higher; this confirms diagnostic utility of this gene as described earlier [15]. Expression of IL2RB is also higher, as seen in Supporting Information S4.…”

Section: Discussionsupporting

confidence: 83%

“…For example, an IL-2 receptor A (IL2RA) enzyme-linked immunosorbent assay (ELISA) exhibited a reported sensitivity of 94% and specificity of 98% [15]. ELISA-based and Griess reaction assays were used to determine that changes in TNF- α and iNOS expression in PBMCs exposed to PPDa or PPDb antigen could serve as additional diagnostic indices complementing IFN-γ measurements [16].…”

Section: Introductionmentioning

confidence: 99%

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Accurate Diagnostics for Bovine tuberculosis Based on High-Throughput Sequencing

Churbanov

Milligan

2012

PLoS ONE

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BackgroundBovine tuberculosis (bTB) is an enduring contagious disease of cattle that has caused substantial losses to the global livestock industry. Despite large-scale eradication efforts, bTB continues to persist. Current bTB tests rely on the measurement of immune responses in vivo (skin tests), and in vitro (bovine interferon-γ release assay). Recent developments are characterized by interrogating the expression of an increasing number of genes that participate in the immune response. Currently used assays have the disadvantages of limited sensitivity and specificity, which may lead to incomplete eradication of bTB. Moreover, bTB that reemerges from wild disease reservoirs requires early and reliable diagnostics to prevent further spread. In this work, we use high-throughput sequencing of the peripheral blood mononuclear cells (PBMCs) transcriptome to identify an extensive panel of genes that participate in the immune response. We also investigate the possibility of developing a reliable bTB classification framework based on RNA-Seq reads.Methodology/Principal FindingsPooled PBMC mRNA samples from unaffected calves as well as from those with disease progression of 1 and 2 months were sequenced using the Illumina Genome Analyzer II. More than 90 million reads were splice-aligned against the reference genome, and deposited to the database for further expression analysis and visualization. Using this database, we identified 2,312 genes that were differentially expressed in response to bTB infection (p<10−8). We achieved a bTB infected status classification accuracy of more than 99% with split-sample validation on newly designed and learned mixtures of expression profiles.Conclusions/SignificanceWe demonstrated that bTB can be accurately diagnosed at the early stages of disease progression based on RNA-Seq high-throughput sequencing. The inclusion of multiple genes in the diagnostic panel, combined with the superior sensitivity and broader dynamic range of RNA-Seq, has the potential to improve the accuracy of bTB diagnostics. The computational pipeline used for the project is available from http://code.google.com/p/bovine-tb-prediction.

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Section: Discussionsupporting

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Accurate Diagnostics for Bovine tuberculosis Based on High-Throughput Sequencing

Churbanov

Milligan

2012

PLoS ONE

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“…Thus, surrogate readout systems of proliferation amenable to whole-blood-based ELISAs are desirable. Determination of production of soluble IL-2 receptors has been used recently with good results (24,25). In this context, our results demonstrating that the levels of IL-2 in 24-h wholeblood culture supernatants are in concordance with the results observed for the LTAs are highly encouraging.…”

Section: Discussionsupporting

confidence: 79%

Development of Diagnostic Reagents To Differentiate betweenMycobacterium bovisBCG Vaccination andM. bovisInfection in Cattle

Vordermeier¹,

Cockle²,

Whelan³

et al. 1999

Clin Diagn Lab Immunol

135

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In Great Britain a recent independent scientific review for the government has concluded that the development of a cattle vaccine against Mycobacterium bovis holds the best long-term prospect for tuberculosis control in British herds. A sine qua non for vaccination is the development of a complementary diagnostic test to differentiate between vaccinated animals and those infected withM. bovis so that test-and-slaughter-based control strategies can continue alongside vaccination. In order to assess the feasibility of developing a differential diagnostic test for a live vaccine, we chose M. bovis BCG Pasteur as a model system. Recombinant forms of antigens which are expressed in M. bovis but not, or only at low levels, in BCG Pasteur (ESAT-6, MPB64, MPB70, and MPB83) were produced. These reagents were tested either alone or in combination by using peripheral blood mononuclear cells from M. bovis-infected, BCG-vaccinated, andMycobacterium avium-sensitized calves. All four antigens induced in vitro proliferation and gamma interferon responses only inM. bovis-infected animals. A cocktail composed of ESAT-6, MPB64, and MPB83 identified infected animals but not those vaccinated with BCG. In addition, promiscuous T-cell epitopes of ESAT-6, MPB64, and MPB83 were formulated into a peptide cocktail. In T-cell assays with this peptide cocktail, infected animals were identified with frequencies similar to those obtained in assays with the protein cocktail, while BCG-vaccinated or M. avium-sensitized animals did not respond. In summary, our results suggest that peptide and protein cocktails can be designed to discriminate between M. bovis infection and BCG vaccination.

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“…A number of strategies have been employed against bovine tb , but the approach has generally been based on government‐organised programmes by which animals deemed positive to defined screening tests are slaughtered. Delayed‐type hypersensitivity ( dth ) skin testing has been the most common test used in cattle (Monaghan and others 1994), but alternative in vitro assays of cellular immunity including lymphocyte proliferation, the release of interferon‐gamma ( ifn ‐γ) and the production of soluble interleukin‐2R ( il ‐2 r ) have also been developed (Outteridge and Lepper 1973, Wood and others 1991, O'Nualláin and others 1997). However, investigations have shown that both the in vivo and the in vitro tests are not 100 per cent specific (Francis and others 1978, Wood and others 1991, Neill and others 1994b, Monaghan and others 1997), and the detection of false‐positives has been a significant part of the economic cost described above, particularly in areas where there is a relatively low incidence of the disease.…”

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confidence: 99%

Assessment of defined antigens for the diagnosis of bovine tuberculosis in skin test‐reactor cattle

Pollock¹,

Rm²,

Ka³

et al. 2000

Veterinary Record

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The continued use of purified protein derivative (PPD) tuberculin is considered to be the main factor which limits the specificity of diagnostic tests for bovine tuberculosis (TB). This study evaluated a whole blood interferon-gamma (IFN-gamma) assay and compared the diagnostic potential of PPD with two tuberculosis-specific antigens, ESAT-6 and MPB70. To provide estimates of sensitivity and specificity, responses were measured in 180 skin test-reacting cattle, of which 131 were confirmed as tuberculous, and in 128 cattle from TB-free herds. For the skin test reactors, there was a positive correlation between the IFN-gamma responses to PPD from Mycobacterium bovis (PPDB) and PPD from Mycobacterium avium (PPDA), indicating cross-reactivity between these complex antigens which are the basis of the skin test. In comparisons of the ESAT-6 IFN-gamma test with a PPD IFN-gamma test (using PPDB compared with PPDA), there was a decrease in sensitivity (76.3 per cent vs 89.3 per cent), but a clear increase in specificity (99.2 per cent vs 92.2 per cent). The provision of high specificity, even with lower sensitivity, offers major benefits for testing in areas with a low incidence of TB.

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Detection of Mycobacterium bovis infection in cattle using an immunoassay for bovine soluble interleukin-2 receptor-α (sIL-2R-α) produced by peripheral blood T-lymphocytes following incubation with tuberculin PPD

Cited by 13 publications

References 20 publications

Accurate Diagnostics for Bovine tuberculosis Based on High-Throughput Sequencing

Accurate Diagnostics for Bovine tuberculosis Based on High-Throughput Sequencing

Development of Diagnostic Reagents To Differentiate betweenMycobacterium bovisBCG Vaccination andM. bovisInfection in Cattle

Assessment of defined antigens for the diagnosis of bovine tuberculosis in skin test‐reactor cattle

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