A reverse transcription-quantitative competitive polymerase chain reaction (RT-qcPCR) technique to measure cytokine gene expression in domestic mammals

被引:56
作者
Rottman, JB [1 ]
Tompkins, WAF [1 ]
Tompkins, MB [1 ]
机构
[1] N CAROLINA STATE UNIV,COLL VET MED,DEPT MICROBIOL PATHOL & PARASITOL,RALEIGH,NC 27606
关键词
cytokines; mRNA expression; quantitative competitive polymerase chain reaction;
D O I
10.1177/030098589603300217
中图分类号
R36 [病理学];
学科分类号
100104 ;
摘要
Inbred strains of rats and mice have long been used to study basic mechanisms of human disease. Our knowledge of the rodent and human immune systems has increased in recent years, largely because of the availability of reagents and techniques specific for these species. In contrast, outbred animals, including domestic companion and food animals, have not been used routinely as experimental models for human disease, largely because reagents and assays necessary for basic research in immunology and physiology have not been available. Here, using consensus cytokine nucleic acid sequences, we adapt a previously described reverse transcription-quantitative competitive polymerase chain reaction technique to measure interleukin 2 (IL2), IL4, IL6, IL10, IL12, interferon gamma, tumor necrosis factor alpha, and glyceraldehyde-3-phosphate dehydrogenase mRNA expression in the cow, cat, dog, horse, and pig. We demonstrate that the assay is sensitive, accurate, and reproducible.
引用
收藏
页码:242 / 248
页数:7
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