Janetzki Sylvia, Cox Josephine H, Oden Neal, Ferrari Guido
ZellNet Consulting Inc., Fort Lee, NJ, USA.
Methods Mol Biol. 2005;302:51-86. doi: 10.1385/1-59259-903-6:051.
During the last 20 yr, the enzyme-linked immunospot (ELISPOT) assay has emerged as one of the most important and widely used assays to monitor immune responses in humans and a variety of other species. With the ELISPOT assay, immune cell frequencies can be measured at the single cell level without elaborate expansion or manipulation of cell populations. Its usefulness has led to its application in vaccine design and development and, most importantly, in monitoring vaccination efforts. The impact of results measured with this assay can be profound. In addition to ease of performance, repeatability and reliability are major features expected of an ELISPOT assay. The focus today is on standardization of the technique, validation strategies to comply with these required features, and accommodation of the growing demand of Good Laboratory Practice (GLP) compliance. This chapter will give the experienced scientists as well as newcomers to the field an overview over the major standardization issues for each step of the protocol. Guidelines are given on how to validate the ELISPOT performance.
在过去20年中，酶联免疫斑点（ELISPOT）检测已成为监测人类和多种其他物种免疫反应的最重要且应用最广泛的检测方法之一。通过ELISPOT检测，可以在单细胞水平测量免疫细胞频率，而无需对细胞群体进行复杂的扩增或处理。其实用性使其应用于疫苗设计与开发，最重要的是，用于监测疫苗接种工作。该检测所测结果的影响可能意义深远。除了操作简便外，重复性和可靠性是ELISPOT检测预期的主要特性。如今的重点在于该技术的标准化、符合这些所需特性的验证策略以及满足对良好实验室规范（GLP）合规性日益增长的需求。本章将为该领域的经验丰富的科学家以及新手提供该方案每个步骤的主要标准化问题概述。文中给出了如何验证ELISPOT性能的指导方针。
