Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum

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dc.contributor.authorCashmore, Tamaryn Jen_ZA
dc.contributor.authorPeter, Jonathan Gen_ZA
dc.contributor.authorVan Zyl-Smit, Richard Nen_ZA
dc.contributor.authorSemple, Patricia Len_ZA
dc.contributor.authorMaredza, Aliceen_ZA
dc.contributor.authorMeldau, Richarden_ZA
dc.contributor.authorZumla, Alimuddinen_ZA
dc.contributor.authorNurse, Barbaraen_ZA
dc.contributor.authorDheda, Keertanen_ZA
dc.date.accessioned2016-01-11T06:51:35Z
dc.date.available2016-01-11T06:51:35Z
dc.date.issued2010en_ZA
dc.description.abstractBACKGROUND: The diagnosis of smear-negative or sputum-scarce tuberculosis (TB) is problematic as culture takes several weeks and representative biological samples are difficult to obtain. RD-1 antigen-specific interferon-γ release assays (IGRAs) are sensitive and specific blood-based tests for the diagnosis of M. tuberculosis infection. The feasibility and diagnostic utility of this rapid immunodiagnostic assay, using cells from induced sputum, is unknown. METHODOLOGY/PRINCIPAL FINDINGS: Cells isolated from induced sputum were co-cultured with ESAT-6 and CFP-10 antigens using a standardized enzyme-linked immunospot (ELISPOT) assay (T-SPOT®. TB ) in 101 consecutively recruited TB suspects or non-TB controls. An optimization phase using 28 samples was followed by a validation phase using samples from 73 participants (20 with definite or probable TB, and 48 with non-TB). Despite optimization of sputum processing 65/73 (89%) of the IGRAs in the validation phase were inconclusive. 44/73 (60%) tests failed due to sputum induction-related factors [sputum induction-related adverse events (n = 5), inadequate sputum volume (n = 8), non-homogenisable sputum (n = 7), and insufficient numbers of cells to perform the assay (n = 24)], whilst 20/73 (27%) tests failed due T-SPOT®. TB assay-related factors [excessive debris precluding reading of spots in the ELISPOT well (n = 6), failure of the positive control (n = 11), or high spot count in the negative control (n = 3)]. Only 8/73 (11%) of the available samples could therefore be correctly categorized (7 definite or probable TB, and 1 non-TB patient). Thus, 13/20 (65%) of the definite or probable TB cases remained undiagnosed. Conclusions/Significance: Rapid immunodiagnosis of pulmonary TB by antigen-specific IFN-γ ELISPOT responses, using cells from induced sputum, is possible. However, the test, in its current ELISPOT format, is not clinically useful because the majority of the assays are inconclusive.en_ZA
dc.identifier.apacitationCashmore, T. J., Peter, J. G., Van Zyl-Smit, R. N., Semple, P. L., Maredza, A., Meldau, R., ... Dheda, K. (2010). Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum. <i>PLoS One</i>, http://hdl.handle.net/11427/16247en_ZA
dc.identifier.chicagocitationCashmore, Tamaryn J, Jonathan G Peter, Richard N Van Zyl-Smit, Patricia L Semple, Alice Maredza, Richard Meldau, Alimuddin Zumla, Barbara Nurse, and Keertan Dheda "Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum." <i>PLoS One</i> (2010) http://hdl.handle.net/11427/16247en_ZA
dc.identifier.citationCashmore, T. J., Peter, J. G., van Zyl-Smit, R. N., Semple, P. L., Maredza, A., Meldau, R., ... & Dheda, K. (2010). Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum. PLoS One, 5(4), e10389. doi:10.1371/journal.pone.0010389en_ZA
dc.identifier.ris TY - Journal Article AU - Cashmore, Tamaryn J AU - Peter, Jonathan G AU - Van Zyl-Smit, Richard N AU - Semple, Patricia L AU - Maredza, Alice AU - Meldau, Richard AU - Zumla, Alimuddin AU - Nurse, Barbara AU - Dheda, Keertan AB - BACKGROUND: The diagnosis of smear-negative or sputum-scarce tuberculosis (TB) is problematic as culture takes several weeks and representative biological samples are difficult to obtain. RD-1 antigen-specific interferon-γ release assays (IGRAs) are sensitive and specific blood-based tests for the diagnosis of M. tuberculosis infection. The feasibility and diagnostic utility of this rapid immunodiagnostic assay, using cells from induced sputum, is unknown. METHODOLOGY/PRINCIPAL FINDINGS: Cells isolated from induced sputum were co-cultured with ESAT-6 and CFP-10 antigens using a standardized enzyme-linked immunospot (ELISPOT) assay (T-SPOT®. TB ) in 101 consecutively recruited TB suspects or non-TB controls. An optimization phase using 28 samples was followed by a validation phase using samples from 73 participants (20 with definite or probable TB, and 48 with non-TB). Despite optimization of sputum processing 65/73 (89%) of the IGRAs in the validation phase were inconclusive. 44/73 (60%) tests failed due to sputum induction-related factors [sputum induction-related adverse events (n = 5), inadequate sputum volume (n = 8), non-homogenisable sputum (n = 7), and insufficient numbers of cells to perform the assay (n = 24)], whilst 20/73 (27%) tests failed due T-SPOT®. TB assay-related factors [excessive debris precluding reading of spots in the ELISPOT well (n = 6), failure of the positive control (n = 11), or high spot count in the negative control (n = 3)]. Only 8/73 (11%) of the available samples could therefore be correctly categorized (7 definite or probable TB, and 1 non-TB patient). Thus, 13/20 (65%) of the definite or probable TB cases remained undiagnosed. Conclusions/Significance: Rapid immunodiagnosis of pulmonary TB by antigen-specific IFN-γ ELISPOT responses, using cells from induced sputum, is possible. However, the test, in its current ELISPOT format, is not clinically useful because the majority of the assays are inconclusive. DA - 2010 DB - OpenUCT DO - 10.1371/journal.pone.0010389 DP - University of Cape Town J1 - PLoS One LK - https://open.uct.ac.za PB - University of Cape Town PY - 2010 T1 - Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum TI - Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum UR - http://hdl.handle.net/11427/16247 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/16247
dc.identifier.urihttp://dx.doi.org/10.1371/journal.pone.0010389
dc.identifier.vancouvercitationCashmore TJ, Peter JG, Van Zyl-Smit RN, Semple PL, Maredza A, Meldau R, et al. Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum. PLoS One. 2010; http://hdl.handle.net/11427/16247.en_ZA
dc.language.isoengen_ZA
dc.publisherPublic Library of Scienceen_ZA
dc.publisher.departmentDivision of Pulmonologyen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.rightsThis is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_ZA
dc.rights.holder© 2010 Cashmore et alen_ZA
dc.rights.urihttp://creativecommons.org/licenses/by/4.0en_ZA
dc.sourcePLoS Oneen_ZA
dc.source.urihttp://journals.plos.org/plosoneen_ZA
dc.subject.otherSputumen_ZA
dc.subject.otherTuberculosisen_ZA
dc.subject.otherEnzyme-linked immunoassaysen_ZA
dc.subject.otherTuberculosis diagnosis and managementen_ZA
dc.subject.otherOptimizationen_ZA
dc.subject.otherDiagnostic medicineen_ZA
dc.subject.otherHIVen_ZA
dc.subject.otherMycobacterium tuberculosisen_ZA
dc.titleFeasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputumen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
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