US Geological Survey, Western Ecological Research Center, Sacramento, CA 95826, USA.
Mol Ecol Resour. 2012 Jan;12(1):67-74. doi: 10.1111/j.1755-0998.2011.03060.x. Epub 2011 Aug 17.
Gene transcription analysis for diagnosing or monitoring wildlife health requires the ability to distinguish pathophysiological change from natural variation. Herein, we describe methodology for the development of quantitative real-time polymerase chain reaction (qPCR) assays to measure differential transcript levels of multiple immune function genes in the sea otter (Enhydra lutris); sea otter-specific qPCR primer sequences for the genes of interest are defined. We establish a 'reference' range of transcripts for each gene in a group of clinically healthy captive and free-ranging sea otters. The 10 genes of interest represent multiple physiological systems that play a role in immuno-modulation, inflammation, cell protection, tumour suppression, cellular stress response, xenobiotic metabolizing enzymes, antioxidant enzymes and cell-cell adhesion. The cycle threshold (C(T)) measures for most genes were normally distributed; the complement cytolysis inhibitor was the exception. The relative enumeration of multiple gene transcripts in simple peripheral blood samples expands the diagnostic capability currently available to assess the health of sea otters in situ and provides a better understanding of the state of their environment.
基因转录分析用于诊断或监测野生动物健康状况，需要能够区分病理生理学变化与自然变异。在此，我们描述了一种开发定量实时聚合酶链反应（qPCR）测定法的方法，用于测量海獭（Enhydra lutris）中多个免疫功能基因的差异转录水平；定义了感兴趣基因的海獭特异性 qPCR 引物序列。我们为一组临床健康的圈养和自由放养的海獭中的每个基因建立了“参考”转录本范围。10 个感兴趣的基因代表多个在免疫调节、炎症、细胞保护、肿瘤抑制、细胞应激反应、外源生物代谢酶、抗氧化酶和细胞-细胞黏附中发挥作用的生理系统。大多数基因的循环阈值（C(T)）测量值呈正态分布；补体细胞溶解抑制剂是个例外。简单外周血样中多个基因转录本的相对计数扩展了目前用于评估海獭现场健康状况的诊断能力，并更好地了解其环境状况。
