4 results on '"Harriet N. Garlant"'
Search Results
2. Evaluation of Host Protein Biomarkers by ELISA From Whole Lysed Peripheral Blood for Development of Diagnostic Tests for Active Tuberculosis
- Author
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Harriet N. Garlant, Kalaiarasan Ellappan, Matthew Hewitt, Prem Perumal, Simon Pekeleke, Nadina Wand, Jo Southern, Saka Vinod Kumar, Harish Belgode, Ibrahim Abubakar, Sanjeev Sinha, Seshadri Vasan, Noyal Mariya Joseph, and Karen E. Kempsell
- Subjects
biomarker ,protein ,ELISA ,tuberculosis ,diagnostic ,assay ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Tuberculosis (TB) remains a significant global health crisis and the number one cause of death for an infectious disease. The health consequences in high-burden countries are significant. Barriers to TB control and eradication are in part caused by difficulties in diagnosis. Improvements in diagnosis are required for organisations like the World Health Organisation (WHO) to meet their ambitious target of reducing the incidence of TB by 50% by the year 2025, which has become hard to reach due to the COVID-19 pandemic. Development of new tests for TB are key priorities of the WHO, as defined in their 2014 report for target product profiles (TPPs). Rapid triage and biomarker-based confirmatory tests would greatly enhance the diagnostic capability for identifying and diagnosing TB-infected individuals. Protein-based test methods e.g. lateral flow devices (LFDs) have a significant advantage over other technologies with regard to assay turnaround time (minutes as opposed to hours) field-ability, ease of use by relatively untrained staff and without the need for supporting laboratory infrastructure. Here we evaluate the diagnostic performance of nine biomarkers from our previously published biomarker qPCR validation study; CALCOCO2, CD274, CD52, GBP1, IFIT3, IFITM3, SAMD9L, SNX10 and TMEM49, as protein targets assayed by ELISA. This preliminary evaluation study was conducted to quantify the level of biomarker protein expression across latent, extra-pulmonary or pulmonary TB groups and negative controls, collected across the UK and India, in whole lysed blood samples (WLB). We also investigated associative correlations between the biomarkers and assessed their suitability for ongoing diagnostic test development, using receiver operating characteristic/area under the curve (ROC) analyses, singly and in panel combinations. The top performing single biomarkers for pulmonary TB versus controls were CALCOCO2, SAMD9L, GBP1, IFITM3, IFIT3 and SNX10. TMEM49 was also significantly differentially expressed but downregulated in TB groups. CD52 expression was not highly differentially expressed across most of the groups but may provide additional patient stratification information and some limited use for incipient latent TB infection. These show therefore great potential for diagnostic test development either in minimal configuration panels for rapid triage or more complex formulations to capture the diversity of disease presentations.
- Published
- 2022
- Full Text
- View/download PDF
3. Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR
- Author
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Prem Perumal, Mohamed Bilal Abdullatif, Harriet N. Garlant, Isobella Honeyborne, Marc Lipman, Timothy D. McHugh, Jo Southern, Ronan Breen, George Santis, Kalaiarasan Ellappan, Saka Vinod Kumar, Harish Belgode, Ibrahim Abubakar, Sanjeev Sinha, Seshadri S. Vasan, Noyal Joseph, and Karen E. Kempsell
- Subjects
tuberculosis ,biomarker ,qPCR ,validation ,diagnosis ,immune ,Immunologic diseases. Allergy ,RC581-607 - Abstract
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).
- Published
- 2021
- Full Text
- View/download PDF
4. Validation of Differentially Expressed Immune Biomarkers in Latent and Active Tuberculosis by Real-Time PCR
- Author
-
Marc Lipman, Timothy D. McHugh, Karen E. Kempsell, Mohamed Bilal Abdullatif, Prem Perumal, Isobella Honeyborne, Harriet N. Garlant, Seshadri S. Vasan, Ronan Breen, Sanjeev Sinha, Harish Narasimha Belgode, George Santis, Jo Southern, Noyal Mariya Joseph, Ibrahim Abubakar, Saka Vinod Kumar, and Kalaiarasan Ellappan
- Subjects
lcsh:Immunologic diseases. Allergy ,0301 basic medicine ,Oncology ,Male ,medicine.medical_specialty ,Tuberculosis ,Asia ,Adolescent ,diagnosis ,Immunology ,Molecular Diagnostic Method ,Real-Time Polymerase Chain Reaction ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Latent Tuberculosis ,Internal medicine ,medicine ,Immunology and Allergy ,Humans ,030212 general & internal medicine ,RNA, Messenger ,Original Research ,validation ,GeneXpert MTB/RIF ,Receiver operating characteristic ,business.industry ,Diagnostic Tests, Routine ,Tuberculin Test ,Incidence (epidemiology) ,Computational Biology ,medicine.disease ,United Kingdom ,qPCR ,030104 developmental biology ,Real-time polymerase chain reaction ,tuberculosis ,ROC Curve ,Biomarker (medicine) ,biomarker ,Female ,immune ,lcsh:RC581-607 ,business ,Biomarkers - Abstract
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).
- Published
- 2021
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