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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Autores principales:	Yosita Panraksa, Anita G Amin, Barbara Graham, Charles S Henry, Delphi Chatterjee
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6904867aefe4450c90ef10b328e8a7c4
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spelling	oai:doaj.org-article:6904867aefe4450c90ef10b328e8a7c42021-12-02T20:14:18ZImmobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis.1932-620310.1371/journal.pone.0257615https://doaj.org/article/6904867aefe4450c90ef10b328e8a7c42021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0257615https://doaj.org/toc/1932-6203The 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.Yosita PanraksaAnita G AminBarbara GrahamCharles S HenryDelphi ChatterjeePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 9, p e0257615 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yosita Panraksa Anita G Amin Barbara Graham Charles S Henry Delphi Chatterjee Immobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis.
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.
format	article
author	Yosita Panraksa Anita G Amin Barbara Graham Charles S Henry Delphi Chatterjee
author_facet	Yosita Panraksa Anita G Amin Barbara Graham Charles S Henry Delphi Chatterjee
author_sort	Yosita Panraksa
title	Immobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis.
title_short	Immobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis.
title_full	Immobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis.
title_fullStr	Immobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis.
title_full_unstemmed	Immobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis.
title_sort	immobilization of proteinase k for urine pretreatment to improve diagnostic accuracy of active tuberculosis.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/6904867aefe4450c90ef10b328e8a7c4
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Immobilization of Proteinase K for urine pretreatment to improve diagnostic accuracy of active tuberculosis.

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