A user-friendly, highly sensitive assay to detect the IFN-γ secretion by T cells
A user-friendly, highly sensitive assay to detect the IFN-γ secretion by T cells
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DOI：
10.1016/j.clinbiochem.2007.12.015
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发表时间：
2008-04-01
影响因子：
2.8
通讯作者：
Geluk, Annemiek
中科院分区：
文献类型：
--
作者：
Corstjens, Paul L. A. M.;Zuiderwijk, Michel;Geluk, Annemiek
目的：开发一个用户友好的测试替代ELISA为基础的测定，以检测IFN-γ在体外培养的外周血单核细胞（PBMC）刺激病原体衍生的antigens.Design and Methods：的分子组成部分的操作IFN-γ ELISA为基础的测试中应用的侧流（LF）免疫夹心测定使用上转换磷（UCP）报告颗粒。的UCP-LF IFN-γ测定（ULIGA）的分析灵敏度进行了测定和定性验证与选择的60个来自PBMC培养物的上清液刺激与麻风杆菌衍生抗原，促分裂原或mediumalone.Results：ULIGA表示分析灵敏度优于2 pg/mL，并表现出四个数量级的动态范围。该测定与IFN-γ ELISA.Conclusions相关性良好：ULIGA允许检测远低于用于定义IFN-γ ELISA中的阳性反应的截止值（100 pg/mL）。该测试程序对设备和劳动力的要求较低，适用于测试单个样品。(c)2007年加拿大临床化学家协会。由爱思唯尔公司出版All rights reserved.
Objectives: Development of a user-friendly test alternative to ELISA-based assays to detect IFN-gamma by in vitro cultured peripheral blood mononuclear cells (PBMC) stimulated with pathogen-derived antigens.Design and methods: The molecular components of an operational IFN-gamma ELISA-based test were applied in a lateral flow (LF) immuno-sandwich assay using up-converting phosphor (UCP) reporter particles. The analytical sensitivity of the UCP-LF IFN-gamma assay (ULIGA) was determined and the assay was qualitatively validated with a selection of 60 supernatants derived from PBMC cultures stimulated with M leprae derived antigens, mitogen or medium alone.Results: ULIGA indicated an analytical sensitivity better than 2 pg/mL, and demonstrated four orders of magnitude dynamic range. The assay correlated well with the IFN-gamma ELISA.Conclusions: ULIGA allows detection well below the cutoff value (100 pg/mL) used to define positive responses in the IFN-gamma ELISA. The test procedure is less demanding in respect to equipment and labor, and is suited for testing single samples. (c) 2007 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.