Accuracy and reproducibility of a multiplex immunoassay platform: A validation study
Accuracy and reproducibility of a multiplex immunoassay platform: A validation study
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DOI：
10.1016/j.jim.2011.01.005
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发表时间：
2011-03-31
影响因子：
2.2
通讯作者：
Ware, Lorraine B.
中科院分区：
文献类型：
--
作者：
Bastarache, Julie A.;Koyama, Tatsuki;Ware, Lorraine B.
Background: Multiplex immunoassays offer many advantages over singleplex assays for the analysis of multiple analytes in a single sample. We sought to validate a specific multiplex cytokine immunoassay (Human 9-plex cytokine array on the Searchlight (R) platform by Thermoscientific) prior to use in a large clinical study.Methods: We compared spike and recovery of recombinant proteins on the Searchlight (R) platform to singleplex immunoassays purchased from R&D Systems, measured identical patient samples on the two different platforms, and measured identical patient samples on different days to measure intra-and inter-assay variability.Results: Assays using the Searchlight (R) platform had inefficient recovery of spiked recombinant proteins compared to R&D Systems singleplex assays. Assaying identical patients samples on different days on the Searchlight platform had acceptable intra-assay variability (intra-assay coefficient of variation (CV%) range for all analytes of 9.1-13.7) but unacceptably high inter-assay variability (CV% range for all analytes 16.7-119.3) suggesting plate-to plate variability. Similar assays for individual cytokines on the R&D platform had an intra-assay CV% range of 1.6-6.4 and an inter-assay CV% range of 3.8-7.1. Some deficiencies in Searchlight (R) assay performance may be due to irregularities in spotting of capture antibodies during manufacturing.Conclusions: We conclude that the Searchlight (R) multiplex immunoassay platform would require extensive additional assay optimization prior to widespread clinical research use. Published by Elsevier B.V.
