Ultra-sensitive urinary lipoarabinomannan (LAM) immunoassay for tuberculosis detection: a performance evaluation.

IF 10.8 1区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

EBioMedicine Pub Date : 2025-09-01 Epub Date: 2025-08-12 DOI:10.1016/j.ebiom.2025.105885

Qisheng Jiang, Carolyn Duncan, Harisha Ramachandraiah, Ige A George, Sumanth Gandra, Marcos Perez, Lorraine Lillis, David S Boyle, Scott Crick, Morten Ruhwald, Srikanth Singamaneni

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引用次数: 0

Abstract

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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超灵敏尿脂阿拉伯糖甘露聚糖（LAM）免疫分析法检测肺结核：性能评价。

背景：发展快速非痰液检测仍然是加速结核病（TB）诊断和治疗启动的全球优先事项。世卫组织唯一推荐的快速诊断检测（RDT）——Alere确定TB lipoarabinman聚糖Ag （AlereLAM）的灵敏度不够理想。基于实验室的电化学发光LAM测定（EclLAM）是当前RDT开发的灵敏度基准和尿液LAM检测的金标准。我们评估了超灵敏的等离子体荧光免疫吸附LAM测定（PFLISA-LAM）与痰专家MTB/RIF、痰培养和尿EclLAM的诊断准确性。方法：建立并评价PFLISA-LAM的检测性能。使用储存的尿液样本进行了两个子研究：临床前研究使用337份特征良好的尿液样本进行截断测定，并初步评估PFLISA-LAM与痰Xpert MTB/RIF和培养相比的性能。2. 一项使用77个盲法样本的诊断准确性评估研究，评估PFLISA-LAM与EclLAM和微生物参考标准（MRS、Xpert阳性和/或培养阳性）相比的性能。结果：PFLISA-LAM检测尿液中LAM的检出限（LOD）为0.84±0.9 pg/mL。在临床前研究中，最佳检测截止值确定为1.7 pg/mL。与培养相比，PFLISA-LAM和痰Xpert MTB/RIF的敏感性分别为51%（95%可信区间[CI]: 43%-59%）和62% （95% CI: 53%-70%）。PFLISA-LAM和Xpert MTB/RIF的特异性分别为99% （95% CI: 96%-100%）和100% （95% CI: 100%-100%）。结合PFLISA-LAM和Xpert MTB/RIF检测数据，可以实现76%的灵敏度提高（95% CI: 69%-83%）。在诊断研究中，EclLAM和PFLISA-LAM检测的敏感性分别为42% （95% CI: 27%-59%）和73% （95% CI: 56%-85%）。EclLAM和PFLISA-LAM的特异性分别为95% （95% CI: 85%-99%）和98% （95% CI: 88%-100%）。结论：与EclLAM相比，PFLISA-LAM具有更好的分析和诊断敏感性，可以更好地检测结核阳性病例的尿LAM。PFLISA-LAM检测也证明了在检测尿液LAM方面提高诊断价值的能力，补充了分子检测，提高了诊断结果。资金：我们没有报告进行这项研究的外部资金支持。

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来源期刊

EBioMedicine Biochemistry, Genetics and Molecular Biology-General Biochemistry,Genetics and Molecular Biology

CiteScore

17.70

自引率

0.90%

发文量

579

审稿时长

5 weeks

期刊介绍： eBioMedicine is a comprehensive biomedical research journal that covers a wide range of studies that are relevant to human health. Our focus is on original research that explores the fundamental factors influencing human health and disease, including the discovery of new therapeutic targets and treatments, the identification of biomarkers and diagnostic tools, and the investigation and modification of disease pathways and mechanisms. We welcome studies from any biomedical discipline that contribute to our understanding of disease and aim to improve human health.