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首页> 外文期刊>Cytokine >Quantifying Aotus monkey cytokines by real-time quantitative RT-PCR.
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Quantifying Aotus monkey cytokines by real-time quantitative RT-PCR.

机译：通过实时定量RT-PCR定量检测大猿猴细胞因子。

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摘要

Aotus spp. monkeys are considered the ideal model for studying the progress of malarial infection and the immune response it elicits. We describe the use of a recently developed technique, real-time quantitative RT-PCR, to quantify several Aotus monkey cytokine mRNAs involved in Th1/Th2 responses (IL-4, IL-10, TNF-beta and IFN-gamma). Specific primers were designed for each cytokine and standard curves were constructed using serial dilutions of pDNA containing each target sequence. Results were normalized to GAPDH housekeeping gene expression levels. Standard curves showed high correlation coefficients and were linear over a wide range of copy numbers. Quantification of Aotus samples showed little intra- and inter-experiment variation, thus, the technique has proven to be highly reproducible and sensitive allowing us to detect as little as 25 copies/microl of target DNA. This technique will allow studying Th1 and Th2 cytokine patterns elicited in response to infection for prospectively evaluating the efficacy of malarial vaccines.
机译：ot属猴子被认为是研究疟疾感染及其引发的免疫反应的理想模型。我们描述了使用一种最新开发的技术，实时定量RT-PCR，以量化参与Th1 / Th2反应（IL-4，IL-10，TNF-β和IFN-γ）的几种Aotus猴细胞因子mRNA。针对每种细胞因子设计特异性引物，并使用含有每种靶序列的pDNA连续稀释液构建标准曲线。将结果标准化为GAPDH管家基因表达水平。标准曲线显示出很高的相关系数，并且在很宽的拷贝数范围内呈线性关系。 Aotus样品的定量显示出实验内和实验间的差异很小，因此，该技术已被证明具有很高的重现性和灵敏性，使我们能够检测到低至25拷贝/微升的靶DNA。该技术将允许研究响应感染引起的Th1和Th2细胞因子模式，从而前瞻性评估疟疾疫苗的功效。

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