Sandwich-type homogeneous chemiluminescence immunoassay based on nanoparticle toward detection of Aspergillus galactomannan antigen

免疫分析 半乳甘露聚糖 化学 化学发光 色谱法 曲霉 曲菌病 同种类的 检出限 抗原 化学发光免疫分析 交叉反应性 抗体 微生物学 免疫学 交叉反应 生物化学 医学 物理 多糖 热力学 生物
作者
Yan Guo,Junpu Li
出处
期刊:Talanta [Elsevier]
卷期号:243: 123392-123392 被引量:5
标识
DOI:10.1016/j.talanta.2022.123392
摘要

Aspergillus species continue to be an important cause of life-threatening infection in immunocompromised patients and galactomannan (GM) is a popular biomarker in the diagnosis of invasive aspergillosis (IA). Here we developed and validated an amplified luminescent proximity homogenous assay-linked immunosorbent assay (AlphaLISA) for serum and bronchoalveolar fluid (BALF) GM based on this approach. Technological processes and reaction conditions were optimized. Study assessments included reproducibility, accuracy, stability, and cross reactivity experiments. Method comparisons with the commercial Platelia Aspergillus enzyme immunoassay (Bio-Rad Laboratories) were performed using 201 clinical serum and BALF samples. Under the optimized conditions, the total runtime of the AlphaLISA method was 40 min with simple operation. The percent coefficient variations (CVs%) were lower than 15%, and the recoveries were in the range of 90-110%. Of note, the proposed assay exhibited acceptable stability and did not display cross-reactivity with non-Aspergillus pathogens. Compared with the results from the Platelia kit, there was a satisfied overall qualitative agreement of 97.5% and overall quantitative correlation coefficient of 0.59. In all, we have successfully developed an alternative novel homogeneous nanoparticle-based immunoassay, which has shorter incubation time and easier protocol than the ones of conventional ELISA. It could serve as an alternative to the well-established Platelia assay for measurement of GM in serum and BALF.
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