Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum
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-gamma release assays (IGRAs) are sensitive and specific blood-based tests for the diagnosis...
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| description | 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-gamma 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.
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.
Rapid immunodiagnosis of pulmonary TB by antigen-specific IFN-gamma 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. |
| doi_str_mv | 10.1371/journal.pone.0010389 |
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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.
Rapid immunodiagnosis of pulmonary TB by antigen-specific IFN-gamma 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.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0010389</identifier><identifier>PMID: 20442850</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adult ; Aged ; Antigens ; Assaying ; Biological properties ; Biological samples ; Case-Control Studies ; Cell culture ; Coculture Techniques ; Diagnosis ; Diagnostic systems ; Enzyme-Linked Immunosorbent Assay ; Enzymes ; ESAT-6 antigen ; Feasibility ; Feasibility Studies ; Female ; Humans ; Immunodiagnosis ; Immunologic Tests - methods ; Immunology ; Immunology/Immune Response ; Immunology/Immunomodulation ; Immunology/Leukocyte Activation ; Infections ; Infectious diseases ; Infectious Diseases/Bacterial Infections ; Infectious Diseases/Respiratory Infections ; Interferon ; Interferon-gamma - analysis ; Interferon-gamma - immunology ; Laboratories ; Male ; Medical diagnosis ; Medicine ; Meningitis ; Microscopy ; Middle Aged ; Optimization ; Patients ; Smear ; Spots ; Sputum ; T cell receptors ; Tuberculosis ; Tuberculosis - diagnosis ; Young Adult ; γ-Interferon</subject><ispartof>PloS one, 2010-04, Vol.5 (4), p.e10389-e10389</ispartof><rights>2010 Cashmore et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>Cashmore et al. 2010</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c525t-954023d278c2801e612016c823096f8a25ed51bacec2ef228c633155baff07653</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2861000/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2861000/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,728,781,785,865,886,2103,2929,23868,27926,27927,53793,53795</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20442850$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Hill, Philip Campbell</contributor><creatorcontrib>Cashmore, Tamaryn J</creatorcontrib><creatorcontrib>Peter, Jonathan G</creatorcontrib><creatorcontrib>van Zyl-Smit, Richard N</creatorcontrib><creatorcontrib>Semple, Patricia L</creatorcontrib><creatorcontrib>Maredza, Alice</creatorcontrib><creatorcontrib>Meldau, Richard</creatorcontrib><creatorcontrib>Zumla, Alimuddin</creatorcontrib><creatorcontrib>Nurse, Barbara</creatorcontrib><creatorcontrib>Dheda, Keertan</creatorcontrib><title>Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>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-gamma 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.
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.
Rapid immunodiagnosis of pulmonary TB by antigen-specific IFN-gamma 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.</description><subject>Adult</subject><subject>Aged</subject><subject>Antigens</subject><subject>Assaying</subject><subject>Biological properties</subject><subject>Biological samples</subject><subject>Case-Control Studies</subject><subject>Cell culture</subject><subject>Coculture Techniques</subject><subject>Diagnosis</subject><subject>Diagnostic systems</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Enzymes</subject><subject>ESAT-6 antigen</subject><subject>Feasibility</subject><subject>Feasibility Studies</subject><subject>Female</subject><subject>Humans</subject><subject>Immunodiagnosis</subject><subject>Immunologic Tests - methods</subject><subject>Immunology</subject><subject>Immunology/Immune Response</subject><subject>Immunology/Immunomodulation</subject><subject>Immunology/Leukocyte Activation</subject><subject>Infections</subject><subject>Infectious diseases</subject><subject>Infectious Diseases/Bacterial Infections</subject><subject>Infectious Diseases/Respiratory Infections</subject><subject>Interferon</subject><subject>Interferon-gamma - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Cashmore, Tamaryn J</au><au>Peter, Jonathan G</au><au>van Zyl-Smit, Richard N</au><au>Semple, Patricia L</au><au>Maredza, Alice</au><au>Meldau, Richard</au><au>Zumla, Alimuddin</au><au>Nurse, Barbara</au><au>Dheda, Keertan</au><au>Hill, Philip Campbell</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2010-04-28</date><risdate>2010</risdate><volume>5</volume><issue>4</issue><spage>e10389</spage><epage>e10389</epage><pages>e10389-e10389</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>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-gamma 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.
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.
Rapid immunodiagnosis of pulmonary TB by antigen-specific IFN-gamma 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.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>20442850</pmid><doi>10.1371/journal.pone.0010389</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2010-04, Vol.5 (4), p.e10389-e10389 |
| issn | 1932-6203 1932-6203 |
| language | eng |
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| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Adult Aged Antigens Assaying Biological properties Biological samples Case-Control Studies Cell culture Coculture Techniques Diagnosis Diagnostic systems Enzyme-Linked Immunosorbent Assay Enzymes ESAT-6 antigen Feasibility Feasibility Studies Female Humans Immunodiagnosis Immunologic Tests - methods Immunology Immunology/Immune Response Immunology/Immunomodulation Immunology/Leukocyte Activation Infections Infectious diseases Infectious Diseases/Bacterial Infections Infectious Diseases/Respiratory Infections Interferon Interferon-gamma - analysis Interferon-gamma - immunology Laboratories Male Medical diagnosis Medicine Meningitis Microscopy Middle Aged Optimization Patients Smear Spots Sputum T cell receptors Tuberculosis Tuberculosis - diagnosis Young Adult γ-Interferon |
| title | Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-18T01%3A49%3A16IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Feasibility%20and%20diagnostic%20utility%20of%20antigen-specific%20interferon-gamma%20responses%20for%20rapid%20immunodiagnosis%20of%20tuberculosis%20using%20induced%20sputum&rft.jtitle=PloS%20one&rft.au=Cashmore,%20Tamaryn%20J&rft.date=2010-04-28&rft.volume=5&rft.issue=4&rft.spage=e10389&rft.epage=e10389&rft.pages=e10389-e10389&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0010389&rft_dat=%3Cproquest_plos_%3E733948948%3C/proquest_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1289453420&rft_id=info:pmid/20442850&rft_doaj_id=oai_doaj_org_article_4ae582f9b83a4717835bb1cd96a8672f&rfr_iscdi=true |