Immobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis
The World Health Organization (WHO) calls for the development of a rapid, biomarker-based, non-sputum test capable of detecting all forms of tuberculosis (TB) at the point-of-care to enable immediate treatment initiation. Lipoarabinomannan (LAM) is the only WHO-endorsed TB biomarker that can be dete...
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| description | The World Health Organization (WHO) calls for the development of a rapid, biomarker-based, non-sputum test capable of detecting all forms of tuberculosis (TB) at the point-of-care to enable immediate treatment initiation. Lipoarabinomannan (LAM) is the only WHO-endorsed TB biomarker that can be detected in urine, an easily collected sample matrix. For obtaining optimal sensitivity, we and others have shown that some form of sample pretreatment is necessary to remove background from patient urine samples. A number of systems are paper-based often destined for resource limited settings. Our current work presents incorporation of one such sample pretreatment, proteinase K (ProK) immobilized on paper (IPK) and test its performance in comparison to standard proteinase K (SPK) treatment that involves addition and deactivation at high temperature prior to performing a capture ELISA. Herein, a simple and economical method was developed for using ProK immobilized strips to pretreat urine samples. Simplification and cost reduction of the proposed pretreatment strip were achieved by using Whatman no.1 paper and by minimizing the concentration of ProK (an expensive but necessary reagent) used to pretreat the clinical samples prior to ELISA. To test the applicability of IPK, capture ELISA was carried out on either LAM-spiked urine or the clinical samples after pretreatment with ProK at 400 μg/mL for 30 minutes at room temperature. The optimal conditions and stability of the IPK were tested and validation was performed on a set of 25 previously analyzed archived clinical urine samples with known TB and HIV status. The results of IPK and SPK treated samples were in agreement showing that the urine LAM test currently under development has the potential to reach adult and pediatric patients regardless of HIV status or site of infection, and to facilitate global TB control to improve assay performance and ultimately treatment outcomes. |
| doi_str_mv | 10.1371/journal.pone.0257615 |
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Lipoarabinomannan (LAM) is the only WHO-endorsed TB biomarker that can be detected in urine, an easily collected sample matrix. For obtaining optimal sensitivity, we and others have shown that some form of sample pretreatment is necessary to remove background from patient urine samples. A number of systems are paper-based often destined for resource limited settings. Our current work presents incorporation of one such sample pretreatment, proteinase K (ProK) immobilized on paper (IPK) and test its performance in comparison to standard proteinase K (SPK) treatment that involves addition and deactivation at high temperature prior to performing a capture ELISA. Herein, a simple and economical method was developed for using ProK immobilized strips to pretreat urine samples. Simplification and cost reduction of the proposed pretreatment strip were achieved by using Whatman no.1 paper and by minimizing the concentration of ProK (an expensive but necessary reagent) used to pretreat the clinical samples prior to ELISA. To test the applicability of IPK, capture ELISA was carried out on either LAM-spiked urine or the clinical samples after pretreatment with ProK at 400 μg/mL for 30 minutes at room temperature. The optimal conditions and stability of the IPK were tested and validation was performed on a set of 25 previously analyzed archived clinical urine samples with known TB and HIV status. The results of IPK and SPK treated samples were in agreement showing that the urine LAM test currently under development has the potential to reach adult and pediatric patients regardless of HIV status or site of infection, and to facilitate global TB control to improve assay performance and ultimately treatment outcomes.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0257615</identifier><identifier>PMID: 34547058</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Binding sites ; Biology and Life Sciences ; Biomarkers ; Biomarkers - urine ; Care and treatment ; COVID-19 ; Deactivation ; Diagnosis ; Endopeptidase K ; Endopeptidase K - chemistry ; Endopeptidase K - metabolism ; Enzyme-linked immunosorbent assay ; Enzyme-Linked Immunosorbent Assay - instrumentation ; Enzyme-Linked Immunosorbent Assay - methods ; Enzymes ; Enzymes, Immobilized - chemistry ; Enzymes, Immobilized - metabolism ; Health aspects ; High temperature ; HIV ; Human immunodeficiency virus ; Humans ; Immobilization ; Immobilized enzymes ; Immunology ; Infections ; Laboratories ; Lipopolysaccharides - urine ; Medical diagnosis ; Medicine and Health Sciences ; Observations ; Pathology ; Patients ; Pediatrics ; Pretreatment ; Proteases ; Proteinase ; Reagents ; Research and Analysis Methods ; Risk factors ; Room temperature ; Sputum ; Temperature ; Tuberculosis ; Tuberculosis - diagnosis ; Urine</subject><ispartof>PloS one, 2021-09, Vol.16 (9), p.e0257615-e0257615</ispartof><rights>COPYRIGHT 2021 Public Library of Science</rights><rights>2021 Panraksa et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://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>2021 Panraksa et al 2021 Panraksa et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-449564c923996aadc9ec228c508b5b01e1e7cc1fa239624d30609c061ac739753</citedby><cites>FETCH-LOGICAL-c758t-449564c923996aadc9ec228c508b5b01e1e7cc1fa239624d30609c061ac739753</cites><orcidid>0000-0002-5215-4699</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8454978/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8454978/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34547058$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Hasnain, Seyed Ehtesham</contributor><creatorcontrib>Panraksa, Yosita</creatorcontrib><creatorcontrib>Amin, Anita G</creatorcontrib><creatorcontrib>Graham, Barbara</creatorcontrib><creatorcontrib>Henry, Charles S</creatorcontrib><creatorcontrib>Chatterjee, Delphi</creatorcontrib><title>Immobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>The World Health Organization (WHO) calls for the development of a rapid, biomarker-based, non-sputum test capable of detecting all forms of tuberculosis (TB) at the point-of-care to enable immediate treatment initiation. 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Lipoarabinomannan (LAM) is the only WHO-endorsed TB biomarker that can be detected in urine, an easily collected sample matrix. For obtaining optimal sensitivity, we and others have shown that some form of sample pretreatment is necessary to remove background from patient urine samples. A number of systems are paper-based often destined for resource limited settings. Our current work presents incorporation of one such sample pretreatment, proteinase K (ProK) immobilized on paper (IPK) and test its performance in comparison to standard proteinase K (SPK) treatment that involves addition and deactivation at high temperature prior to performing a capture ELISA. Herein, a simple and economical method was developed for using ProK immobilized strips to pretreat urine samples. Simplification and cost reduction of the proposed pretreatment strip were achieved by using Whatman no.1 paper and by minimizing the concentration of ProK (an expensive but necessary reagent) used to pretreat the clinical samples prior to ELISA. To test the applicability of IPK, capture ELISA was carried out on either LAM-spiked urine or the clinical samples after pretreatment with ProK at 400 μg/mL for 30 minutes at room temperature. The optimal conditions and stability of the IPK were tested and validation was performed on a set of 25 previously analyzed archived clinical urine samples with known TB and HIV status. The results of IPK and SPK treated samples were in agreement showing that the urine LAM test currently under development has the potential to reach adult and pediatric patients regardless of HIV status or site of infection, and to facilitate global TB control to improve assay performance and ultimately treatment outcomes.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>34547058</pmid><doi>10.1371/journal.pone.0257615</doi><tpages>e0257615</tpages><orcidid>https://orcid.org/0000-0002-5215-4699</orcidid><oa>free_for_read</oa></addata></record> |
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| issn | 1932-6203 1932-6203 |
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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 | Binding sites Biology and Life Sciences Biomarkers Biomarkers - urine Care and treatment COVID-19 Deactivation Diagnosis Endopeptidase K Endopeptidase K - chemistry Endopeptidase K - metabolism Enzyme-linked immunosorbent assay Enzyme-Linked Immunosorbent Assay - instrumentation Enzyme-Linked Immunosorbent Assay - methods Enzymes Enzymes, Immobilized - chemistry Enzymes, Immobilized - metabolism Health aspects High temperature HIV Human immunodeficiency virus Humans Immobilization Immobilized enzymes Immunology Infections Laboratories Lipopolysaccharides - urine Medical diagnosis Medicine and Health Sciences Observations Pathology Patients Pediatrics Pretreatment Proteases Proteinase Reagents Research and Analysis Methods Risk factors Room temperature Sputum Temperature Tuberculosis Tuberculosis - diagnosis Urine |
| title | Immobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-20T12%3A31%3A58IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Immobilization%20of%20Proteinase%20K%20for%20urine%20pretreatment%20to%20improve%20diagnostic%20accuracy%20of%20active%20tuberculosis&rft.jtitle=PloS%20one&rft.au=Panraksa,%20Yosita&rft.date=2021-09-21&rft.volume=16&rft.issue=9&rft.spage=e0257615&rft.epage=e0257615&rft.pages=e0257615-e0257615&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0257615&rft_dat=%3Cgale_plos_%3EA676312380%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2575053409&rft_id=info:pmid/34547058&rft_galeid=A676312380&rft_doaj_id=oai_doaj_org_article_6904867aefe4450c90ef10b328e8a7c4&rfr_iscdi=true |