Response definition criteria for ELISPOT assays revisited.
Response definition criteria for ELISPOT assays revisited.
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DOI：
10.1007/s00262-010-0875-4
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发表时间：
2010-10
期刊：
影响因子：
--
通讯作者：
Britten CM
中科院分区：
文献类型：
--
作者：
Moodie Z;Price L;Gouttefangeas C;Mander A;Janetzki S;Löwer M;Welters MJ;Ottensmeier C;van der Burg SH;Britten CM
关于如何根据ELISPOT测定的原始数据确定是否检测到免疫应答，尚未达成共识。本文的目的是使研究人员能够理解并容易地实施当前可用的方法来确定响应。我们描述了经验和统计方法，确定每种方法的优势和局限性，使读者能够合理选择和应用适合其特定实验室设置的科学合理的方法。将五种代表性方法应用于CIMT免疫引导计划的数据集，并比较反应检出率和假阳性率。模拟研究也进行比较经验和统计方法。基于这些，我们建议使用非参数统计检验。此外，我们建议在培养基控制和实验条件下分别进行6个培养基控制井或4个培养基控制井，以提高检测响应的灵敏度，过滤掉斑点计数变化较大的重复，并且谨慎解释实验斑点计数低于估计检测极限的阳性响应。此外，还开发了一个基于网络的用户界面，以便方便地使用所建议的统计方法。该界面允许用户上传ELISPOT分析的数据，并获得二进制响应的输出文件。本文的在线版本（doi:10.1007/s00262-010-0875-4）包含补充材料，可供授权用户使用。
No consensus has been reached on how to determine if an immune response has been detected based on raw data from an ELISPOT assay. The goal of this paper is to enable investigators to understand and readily implement currently available methods for response determination. We describe empirical and statistical approaches, identifying the strengths and limitations of each approach to allow readers to rationally select and apply a scientifically sound method appropriate to their specific laboratory setting. Five representative approaches were applied to data sets from the CIMT Immunoguiding Program and the response detection and false positive rates were compared. Simulation studies were also performed to compare empirical and statistical approaches. Based on these, we recommend the use of a non-parametric statistical test. Further, we recommend that six medium control wells or four wells each for both medium control and experimental conditions be performed to increase the sensitivity in detecting a response, that replicates with large variation in spot counts be filtered out, and that positive responses arising from experimental spot counts below the estimated limit of detection be interpreted with caution. Moreover, a web-based user interface was developed to allow easy access to the recommended statistical methods. This interface allows the user to upload data from an ELISPOT assay and obtain an output file of the binary responses. The online version of this article (doi:10.1007/s00262-010-0875-4) contains supplementary material, which is available to authorized users.
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DOI：
10.1007/s00262-009-0814-4
发表时间：
2010-04
期刊：
Cancer immunology, immunotherapy : CII
影响因子：
--
作者：
Mander A;Gouttefangeas C;Ottensmeier C;Welters MJ;Low L;van der Burg SH;Britten CM
通讯作者：
Britten CM
影响因子：
6.4
作者：
Herr, W;Protzer, U;Wölfel, T
通讯作者：
Wölfel, T
影响因子：
5.8
作者：
Janetzki, S.;Janetzki, S.;Hoos, A.
通讯作者：
Hoos, A.
影响因子：
2
作者：
HOCHBERG, Y;BENJAMINI, Y
通讯作者：
BENJAMINI, Y
影响因子：
2.2
作者：
Moodie, Zoe;Huang, Yunda;Self, Steven G.
通讯作者：
Self, Steven G.