Evaluation of Host Protein Biomarkers by ELISA From Whole Lysed Peripheral Blood for Development of Diagnostic Tests for Active Tuberculosis

Frontiers in Immunology

Seshadri Vasan¹  Saka Vinod Kumar¹  Karen E. Kempsell²  Noyal Mariya Joseph³  Harish Belgode⁴  Kalaiarasan Ellappan⁴  Ibrahim Abubakar⁴  Sanjeev Sinha⁵  Jo Southern⁵  Harriet N. Garlant⁶  Nadina Wand⁶  Prem Perumal⁶  Matthew Hewitt⁶  Simon Pekeleke⁶ 

[1] Medical Sciences, Mortimer Market Centre, University College London, London, United Kingdom;Department of Health Sciences, University of York, York, United Kingdom;Department of Medicine, All India Institute for Medical Sciences, New Delhi, India;Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India;;School of Life &Science Group: Research and Evaluation, UK Health Security Agency, Salisbury, United Kingdom;

关键词: biomarker;    protein;    ELISA;    tuberculosis;    diagnostic;    assay;   

DOI  :  10.3389/fimmu.2022.854327

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown