Interpreting ELISA analyses from wild animal samples: some recurrent issues and solutions

1. Many studies in disease and immunological ecology rely on the use of assays that quantify the amount of specific antibodies (immunoglobulin) in samples. Enzyme-Linked Immuno Sorbent Assays (ELISAs) are increasingly used in ecology due to their availability for a broad array of antigens and the limited amount of sampling material they require. Two recurrent methodological issues are nevertheless faced by researchers: (i) the limited availability of immunological assays and reagents developed for non-model species, and (ii) the statistical determination of the cut-off threshold used to distinguish individual samples that are likely to have or not to have antibodies against a specific antigen.
2. Here, we outline two solutions to deal with these issues. First, we show that implementing two assays with differing detection methods can help validate the use of reagents, such as antibodies, in species different from their intended target. We illustrate this by comparing the quantification ...

1. 诸多疾病与免疫生态学研究均采用可定量检测样本中特异性抗体（免疫球蛋白，immunoglobulin）含量的免疫检测实验。酶联免疫吸附试验（Enzyme-Linked Immuno Sorbent Assays, ELISAs）因可适配多种抗原且所需采样样本量极少，在生态学研究中的应用愈发广泛。但研究者常面临两类常见方法论难题：其一，针对非模式物种开发的免疫检测试剂与方法供应有限；其二，如何通过统计学方法确定临界阈值（cut-off threshold），以区分样本是否含有针对特定抗原的抗体。
2. 本研究针对上述两类难题提出两种解决方案。首先，我们证实采用两种检测原理不同的免疫检测实验，可有效验证试剂（如抗体）在非靶标物种中的适用性。我们通过对比定量分析……