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Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR

Tuberculosis (TB) remains a major global threat and diagnosis of active TB ((ATB) both extra-pulmonary (EPTB), pulmonary (PTB)) and latent TB (LTBI) infection remains challenging, particularly in high-burden countries which still rely heavily on conventional methods. Although molecular diagnostic me...

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Autores principales: Perumal, Prem, Abdullatif, Mohamed Bilal, Garlant, Harriet N., Honeyborne, Isobella, Lipman, Marc, McHugh, Timothy D., Southern, Jo, Breen, Ronan, Santis, George, Ellappan, Kalaiarasan, Kumar, Saka Vinod, Belgode, Harish, Abubakar, Ibrahim, Sinha, Sanjeev, Vasan, Seshadri S., Joseph, Noyal, Kempsell, Karen E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8029985/
https://www.ncbi.nlm.nih.gov/pubmed/33841389
http://dx.doi.org/10.3389/fimmu.2020.612564
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author Perumal, Prem
Abdullatif, Mohamed Bilal
Garlant, Harriet N.
Honeyborne, Isobella
Lipman, Marc
McHugh, Timothy D.
Southern, Jo
Breen, Ronan
Santis, George
Ellappan, Kalaiarasan
Kumar, Saka Vinod
Belgode, Harish
Abubakar, Ibrahim
Sinha, Sanjeev
Vasan, Seshadri S.
Joseph, Noyal
Kempsell, Karen E.
author_facet Perumal, Prem
Abdullatif, Mohamed Bilal
Garlant, Harriet N.
Honeyborne, Isobella
Lipman, Marc
McHugh, Timothy D.
Southern, Jo
Breen, Ronan
Santis, George
Ellappan, Kalaiarasan
Kumar, Saka Vinod
Belgode, Harish
Abubakar, Ibrahim
Sinha, Sanjeev
Vasan, Seshadri S.
Joseph, Noyal
Kempsell, Karen E.
author_sort Perumal, Prem
collection PubMed
description Tuberculosis (TB) remains a major global threat and diagnosis of active TB ((ATB) both extra-pulmonary (EPTB), pulmonary (PTB)) and latent TB (LTBI) infection remains challenging, particularly in high-burden countries which still rely heavily on conventional methods. Although molecular diagnostic methods are available, e.g., Cepheid GeneXpert, they are not universally available in all high TB burden countries. There is intense focus on immune biomarkers for use in TB diagnosis, which could provide alternative low-cost, rapid diagnostic solutions. In our previous gene expression studies, we identified peripheral blood leukocyte (PBL) mRNA biomarkers in a non-human primate TB aerosol-challenge model. Here, we describe a study to further validate select mRNA biomarkers from this prior study in new cohorts of patients and controls, as a prerequisite for further development. Whole blood mRNA was purified from ATB patients recruited in the UK and India, LTBI and two groups of controls from the UK (i) a low TB incidence region (CNTRLA) and (ii) individuals variably-domiciled in the UK and Asia ((CNTRLB), the latter TB high incidence regions). Seventy-two mRNA biomarker gene targets were analyzed by qPCR using the Roche Lightcycler 480 qPCR platform and data analyzed using GeneSpring™ 14.9 bioinformatics software. Differential expression of fifty-three biomarkers was confirmed between MTB infected, LTBI groups and controls, seventeen of which were significant using analysis of variance (ANOVA): CALCOCO2, CD52, GBP1, GBP2, GBP5, HLA-B, IFIT3, IFITM3, IRF1, LOC400759 (GBP1P1), NCF1C, PF4V1, SAMD9L, S100A11, TAF10, TAPBP, and TRIM25. These were analyzed using receiver operating characteristic (ROC) curve analysis. Single biomarkers and biomarker combinations were further assessed using simple arithmetic algorithms. Minimal combination biomarker panels were delineated for primary diagnosis of ATB (both PTB and EPTB), LTBI and identifying LTBI individuals at high risk of progression which showed good performance characteristics. These were assessed for suitability for progression against the standards for new TB diagnostic tests delineated in the published World Health Organization (WHO) technology product profiles (TPPs).
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spelling pubmed-80299852021-04-09 Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR Perumal, Prem Abdullatif, Mohamed Bilal Garlant, Harriet N. Honeyborne, Isobella Lipman, Marc McHugh, Timothy D. Southern, Jo Breen, Ronan Santis, George Ellappan, Kalaiarasan Kumar, Saka Vinod Belgode, Harish Abubakar, Ibrahim Sinha, Sanjeev Vasan, Seshadri S. Joseph, Noyal Kempsell, Karen E. Front Immunol Immunology Tuberculosis (TB) remains a major global threat and diagnosis of active TB ((ATB) both extra-pulmonary (EPTB), pulmonary (PTB)) and latent TB (LTBI) infection remains challenging, particularly in high-burden countries which still rely heavily on conventional methods. Although molecular diagnostic methods are available, e.g., Cepheid GeneXpert, they are not universally available in all high TB burden countries. There is intense focus on immune biomarkers for use in TB diagnosis, which could provide alternative low-cost, rapid diagnostic solutions. In our previous gene expression studies, we identified peripheral blood leukocyte (PBL) mRNA biomarkers in a non-human primate TB aerosol-challenge model. Here, we describe a study to further validate select mRNA biomarkers from this prior study in new cohorts of patients and controls, as a prerequisite for further development. Whole blood mRNA was purified from ATB patients recruited in the UK and India, LTBI and two groups of controls from the UK (i) a low TB incidence region (CNTRLA) and (ii) individuals variably-domiciled in the UK and Asia ((CNTRLB), the latter TB high incidence regions). Seventy-two mRNA biomarker gene targets were analyzed by qPCR using the Roche Lightcycler 480 qPCR platform and data analyzed using GeneSpring™ 14.9 bioinformatics software. Differential expression of fifty-three biomarkers was confirmed between MTB infected, LTBI groups and controls, seventeen of which were significant using analysis of variance (ANOVA): CALCOCO2, CD52, GBP1, GBP2, GBP5, HLA-B, IFIT3, IFITM3, IRF1, LOC400759 (GBP1P1), NCF1C, PF4V1, SAMD9L, S100A11, TAF10, TAPBP, and TRIM25. These were analyzed using receiver operating characteristic (ROC) curve analysis. Single biomarkers and biomarker combinations were further assessed using simple arithmetic algorithms. Minimal combination biomarker panels were delineated for primary diagnosis of ATB (both PTB and EPTB), LTBI and identifying LTBI individuals at high risk of progression which showed good performance characteristics. These were assessed for suitability for progression against the standards for new TB diagnostic tests delineated in the published World Health Organization (WHO) technology product profiles (TPPs). Frontiers Media S.A. 2021-03-16 /pmc/articles/PMC8029985/ /pubmed/33841389 http://dx.doi.org/10.3389/fimmu.2020.612564 Text en Copyright © 2021 Perumal, Abdullatif, Garlant, Honeyborne, Lipman, McHugh, Southern, Breen, Santis, Ellappan, Kumar, Belgode, Abubakar, Sinha, Vasan, Joseph and Kempsell https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Perumal, Prem
Abdullatif, Mohamed Bilal
Garlant, Harriet N.
Honeyborne, Isobella
Lipman, Marc
McHugh, Timothy D.
Southern, Jo
Breen, Ronan
Santis, George
Ellappan, Kalaiarasan
Kumar, Saka Vinod
Belgode, Harish
Abubakar, Ibrahim
Sinha, Sanjeev
Vasan, Seshadri S.
Joseph, Noyal
Kempsell, Karen E.
Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR
title Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR
title_full Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR
title_fullStr Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR
title_full_unstemmed Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR
title_short Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR
title_sort validation of differentially expressed immune biomarkers in latent and active tuberculosis by real-time pcr
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8029985/
https://www.ncbi.nlm.nih.gov/pubmed/33841389
http://dx.doi.org/10.3389/fimmu.2020.612564
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