Ultra-sensitive urinary lipoarabinomannan (LAM) immunoassay for tuberculosis detection: a performance evaluationResearch in context

Summary: Background: The development of rapid non-sputum tests remains a global priority to accelerate Tuberculosis (TB) diagnosis and treatment initiation. The only WHO-recommended rapid diagnostic test (RDT), the Alere Determine TB Lipoarabinomannan Ag (AlereLAM) has suboptimal sensitivity. A lab...

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Main Authors: Qisheng Jiang, Carolyn Duncan, Harisha Ramachandraiah, Ige A. George, Sumanth Gandra, Marcos Perez, Lorraine Lillis, David S. Boyle, Scott Crick, Morten Ruhwald, Srikanth Singamaneni
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:EBioMedicine
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352396425003299
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author Qisheng Jiang
Carolyn Duncan
Harisha Ramachandraiah
Ige A. George
Sumanth Gandra
Marcos Perez
Lorraine Lillis
David S. Boyle
Scott Crick
Morten Ruhwald
Srikanth Singamaneni
author_facet Qisheng Jiang
Carolyn Duncan
Harisha Ramachandraiah
Ige A. George
Sumanth Gandra
Marcos Perez
Lorraine Lillis
David S. Boyle
Scott Crick
Morten Ruhwald
Srikanth Singamaneni
author_sort Qisheng Jiang
collection DOAJ
description Summary: Background: The development of rapid non-sputum tests remains a global priority to accelerate Tuberculosis (TB) diagnosis and treatment initiation. The only WHO-recommended rapid diagnostic test (RDT), the Alere Determine TB Lipoarabinomannan Ag (AlereLAM) has suboptimal sensitivity. A laboratory-based electrochemiluminescence LAM assay (EclLAM) is the current sensitivity benchmark for RDT development and the gold standard for urinary LAM detection. We assessed the diagnostic accuracy of an ultra-sensitive, Plasmonic Fluor-linked Immunosorbent LAM assay (PFLISA-LAM) compared to Sputum Xpert MTB/RIF, sputum culture and urine EclLAM. Methods: We developed and evaluated the assay performance of PFLISA-LAM. Two sub-studies were conducted using banked urine samples: 1. Preclinical study using 337 well-characterised urine samples for cutoff determination and initial evaluation of the performance of PFLISA-LAM compared to sputum Xpert MTB/RIF and culture. 2. A Diagnostic accuracy assessment study using 77 blinded samples to evaluate the performance of PFLISA-LAM compared to EclLAM versus microbiological reference standard (MRS, Xpert positive and/or culture positive). Findings: PFLISA-LAM has a limit of detection (LOD) of 0.84 ± 0.9 pg/mL when detecting purified LAM spiked in urine. In the preclinical study, the optimal assay cutoff was determined to be 1.7 pg/mL. The sensitivities of PFLISA-LAM and sputum Xpert MTB/RIF compared to culture were 51% (95% confidence interval [CI]: 43%–59%) and 62% (95% CI: 53%–70%). The specificities of PFLISA-LAM and Xpert MTB/RIF were 99% (95% CI: 96%–100%) and 100% (95% CI: 100%–100%). Combining PFLISA-LAM and Xpert MTB/RIF test data, an improved sensitivity of 76% (95% CI: 69%–83%) can be achieved. In the diagnostic study, the sensitivities of EclLAM and PFLISA-LAM assays were 42% (95% CI: 27%–59%) and 73% (95% CI: 56%–85%). The specificities of EclLAM and PFLISA-LAM were 95% (95% CI: 85%–99%) and 98% (95% CI: 88%–100%). Interpretation: With better analytical and diagnostic sensitivity compared to EclLAM, PFLISA-LAM can better detect urinary LAM in TB-positive cases. PFLISA-LAM assay also demonstrated the capability to increase the diagnostic value in detecting urinary LAM, complementing molecular tests, achieving improved diagnostic outcome. Funding: We report no external financial support for conducting the study.
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spelling doaj-art-b2b61fe6eae8403b8d8e61ed37b13c9d2025-08-20T03:41:54ZengElsevierEBioMedicine2352-39642025-09-0111910588510.1016/j.ebiom.2025.105885Ultra-sensitive urinary lipoarabinomannan (LAM) immunoassay for tuberculosis detection: a performance evaluationResearch in contextQisheng Jiang0Carolyn Duncan1Harisha Ramachandraiah2Ige A. George3Sumanth Gandra4Marcos Perez5Lorraine Lillis6David S. Boyle7Scott Crick8Morten Ruhwald9Srikanth Singamaneni10Brightest Bio, St. Louis, MO, 63114, United States; Corresponding author.Brightest Bio, St. Louis, MO, 63114, United StatesFIND, The Global Alliance for Diagnostics, Geneva, SwitzerlandDivision of Infectious Diseases, Department of Medicine, Washington University School of Medicine, St. Louis, MO, 63130, United StatesDivision of Infectious Diseases, Department of Medicine, Washington University School of Medicine, St. Louis, MO, 63130, United StatesPATH, Seattle, WA, 98121, United StatesPATH, Seattle, WA, 98121, United StatesPATH, Seattle, WA, 98121, United StatesBrightest Bio, St. Louis, MO, 63114, United StatesFIND, The Global Alliance for Diagnostics, Geneva, SwitzerlandDepartment of Mechanical Engineering and Materials Science, Institute of Materials Science and Engineering, Washington University in St. Louis, St. Louis, MO, 63130, United StatesSummary: Background: The development of rapid non-sputum tests remains a global priority to accelerate Tuberculosis (TB) diagnosis and treatment initiation. The only WHO-recommended rapid diagnostic test (RDT), the Alere Determine TB Lipoarabinomannan Ag (AlereLAM) has suboptimal sensitivity. A laboratory-based electrochemiluminescence LAM assay (EclLAM) is the current sensitivity benchmark for RDT development and the gold standard for urinary LAM detection. We assessed the diagnostic accuracy of an ultra-sensitive, Plasmonic Fluor-linked Immunosorbent LAM assay (PFLISA-LAM) compared to Sputum Xpert MTB/RIF, sputum culture and urine EclLAM. Methods: We developed and evaluated the assay performance of PFLISA-LAM. Two sub-studies were conducted using banked urine samples: 1. Preclinical study using 337 well-characterised urine samples for cutoff determination and initial evaluation of the performance of PFLISA-LAM compared to sputum Xpert MTB/RIF and culture. 2. A Diagnostic accuracy assessment study using 77 blinded samples to evaluate the performance of PFLISA-LAM compared to EclLAM versus microbiological reference standard (MRS, Xpert positive and/or culture positive). Findings: PFLISA-LAM has a limit of detection (LOD) of 0.84 ± 0.9 pg/mL when detecting purified LAM spiked in urine. In the preclinical study, the optimal assay cutoff was determined to be 1.7 pg/mL. The sensitivities of PFLISA-LAM and sputum Xpert MTB/RIF compared to culture were 51% (95% confidence interval [CI]: 43%–59%) and 62% (95% CI: 53%–70%). The specificities of PFLISA-LAM and Xpert MTB/RIF were 99% (95% CI: 96%–100%) and 100% (95% CI: 100%–100%). Combining PFLISA-LAM and Xpert MTB/RIF test data, an improved sensitivity of 76% (95% CI: 69%–83%) can be achieved. In the diagnostic study, the sensitivities of EclLAM and PFLISA-LAM assays were 42% (95% CI: 27%–59%) and 73% (95% CI: 56%–85%). The specificities of EclLAM and PFLISA-LAM were 95% (95% CI: 85%–99%) and 98% (95% CI: 88%–100%). Interpretation: With better analytical and diagnostic sensitivity compared to EclLAM, PFLISA-LAM can better detect urinary LAM in TB-positive cases. PFLISA-LAM assay also demonstrated the capability to increase the diagnostic value in detecting urinary LAM, complementing molecular tests, achieving improved diagnostic outcome. Funding: We report no external financial support for conducting the study.http://www.sciencedirect.com/science/article/pii/S2352396425003299TuberculosisLipoarabinomannanNon-sputumDiagnostic accuracyUltra-sensitive assay
spellingShingle Qisheng Jiang
Carolyn Duncan
Harisha Ramachandraiah
Ige A. George
Sumanth Gandra
Marcos Perez
Lorraine Lillis
David S. Boyle
Scott Crick
Morten Ruhwald
Srikanth Singamaneni
Ultra-sensitive urinary lipoarabinomannan (LAM) immunoassay for tuberculosis detection: a performance evaluationResearch in context
EBioMedicine
Tuberculosis
Lipoarabinomannan
Non-sputum
Diagnostic accuracy
Ultra-sensitive assay
title Ultra-sensitive urinary lipoarabinomannan (LAM) immunoassay for tuberculosis detection: a performance evaluationResearch in context
title_full Ultra-sensitive urinary lipoarabinomannan (LAM) immunoassay for tuberculosis detection: a performance evaluationResearch in context
title_fullStr Ultra-sensitive urinary lipoarabinomannan (LAM) immunoassay for tuberculosis detection: a performance evaluationResearch in context
title_full_unstemmed Ultra-sensitive urinary lipoarabinomannan (LAM) immunoassay for tuberculosis detection: a performance evaluationResearch in context
title_short Ultra-sensitive urinary lipoarabinomannan (LAM) immunoassay for tuberculosis detection: a performance evaluationResearch in context
title_sort ultra sensitive urinary lipoarabinomannan lam immunoassay for tuberculosis detection a performance evaluationresearch in context
topic Tuberculosis
Lipoarabinomannan
Non-sputum
Diagnostic accuracy
Ultra-sensitive assay
url http://www.sciencedirect.com/science/article/pii/S2352396425003299
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