Comparison of Dissociation-Enhanced Lanthanide Fluorescent Immunoassays to Enzyme-Linked Immunosorbent Assays for Detection of Staphylococcal Enterotoxin B, Yersinia pestis-Specific F1 Antigen, and Venezuelan Equine Encephalitis Virus
Comparison of Dissociation-Enhanced Lanthanide Fluorescent Immunoassays to Enzyme-Linked Immunosorbent Assays for Detection of Staphylococcal Enterotoxin B, Yersinia pestis-Specific F1 Antigen, and Venezuelan Equine Encephalitis Virus
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解离增强镧系元素荧光免疫分析与酶联免疫吸附分析检测葡萄球菌肠毒素 B、鼠疫耶尔森氏菌特异性 F1 抗原和委内瑞拉马脑炎病毒的比较
DOI：
10.1128/cdli.8.6.1070-1075.2001
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
G. Ludwig
中科院分区：
文献类型：
--
作者：
Darci R Smith;C. Rossi;T. Kijek;E. Henchal;G. Ludwig
建立了分离增强镧系元素荧光免疫分析法（DELFIA），用于检测葡萄球菌肠毒素B、鼠疫耶尔森氏菌特异性F1抗原和委内瑞拉马脑炎病毒。通过测定灵敏度或检测限（LOD）、动态范围和多种不同样品基质中各检测方法的重现性，将这些检测方法与先前开发的酶联免疫吸附测定（ELISA）进行比较。然后使用一小组设盲加标和未加标样品测定各试验的灵敏度和特异性。所有三种DELFIAs的灵敏度均比使用相同试剂的相应ELISA高至少1个对数，并且动态范围增加至少2个对数10浓度。这种增加的LOD导致DELFIA的灵敏度更高。评价的所有试验的特异性均为100%，在两种形式中均未观察到样品基质效应。然而，由于随机分布的威尔斯孔显示出过度的背景信号（热威尔斯孔），DELFIA的再现性较差，这在整个评价过程中都会发生。随着这项技术的成熟，这些检测的重现性应该会提高，识别热威尔斯的能力也会提高。尽管其敏感性，但与DELFIA相关的后勤负担以及完成测定和解释数据所需的技术专业知识限制了该技术在参考或大型临床实验室的应用。
ABSTRACT The dissociation-enhanced lanthanide fluorescent immunoassays (DELFIA) were developed for the detection of staphylococcal enterotoxin B, Yersinia pestis-specific F1 antigen, and Venezuelan equine encephalitis virus. These assays were compared to previously developed enzyme-linked immunosorbent assays (ELISAs) by determining the sensitivity or limit of detection (LOD), the dynamic range, and the reproducibility of each assay in a number of different sample matrices. The sensitivity and specificity of each assay were then determined by using a small panel of blinded spiked and nonspiked samples. All three DELFIAs demonstrated at least 1 log greater sensitivity than corresponding ELISAs utilizing the same reagents and showed an increase in dynamic range of at least 2 log10 concentrations. This increased LOD resulted in higher sensitivity rates for the DELFIA. The specificity of all of the assays evaluated was 100%, and no sample matrix effects were observed in either format. However, the reproducibility of the DELFIA was poor due to randomly distributed wells exhibiting excessive background signal (hot wells), which occurred throughout the evaluation. As this technology matures, the reproducibility of these assays should improve, as will the ability to identify hot wells. Despite its sensitivity, the logistical burden associated with the DELFIA and the technical expertise required to complete assays and interpret the data limit the application of this technology to reference or large clinical laboratories.