Comparison of RT-qPCR and Nanostring in the measurement of blood interferon response for the diagnosis of type I interferonopathies
Comparison of RT-qPCR and Nanostring in the measurement of blood interferon response for the diagnosis of type I interferonopathies
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DOI：
10.1016/j.cyto.2018.10.023
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发表时间：
2019-01-01
期刊：
影响因子：
3.8
通讯作者：
Viel, Sebastien
中科院分区：
文献类型：
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作者：
Pescarmona, Remi;Belot, Alexandre;Viel, Sebastien
I 型干扰素病的特征是循环 I 型干扰素 (IFN) 浓度增加。 I 型干扰素病是指罕见的孟德尔遗传性疾病，例如艾卡迪-古蒂埃综合征 (AGS) 以及更常见的多基因自身免疫性疾病，例如系统性红斑狼疮 (SLE)。然而，在这些患者中检测 I 型干扰素仍然具有挑战性，因为患者血清中的 I 型干扰素含量通常非常低。因此，已提出检测干扰素刺激基因作为检测该细胞因子的替代方法，但迄今为止尚未报道该测定的灵敏度、特异性和预测值。在本研究中，我们提出了两种基于 Nanostring 或 RT-qPCR 的不同方法，用于在临床常规中测量 IFN 反应，定义为由 IFN 系统诱导的一组转录物。 IFN 特征由 6 个 IFN 刺激基因 (ISG) 组成，对 I 型干扰素病的诊断具有很强的预测价值。这种简单测试的使用可能代表了评估各种自身免疫性疾病的黄金标准。此外，该测试还可用于监测接受针对 I 型 IFN 途径的药物治疗的患者。在比较 Nanostring 和 qPCR 两种方法的分析性能时，它们提供了相似的结果，但 Nanostring 更快、更容易进行多重检测，并且几乎完全自动化，这代表了日常临床实践中更可靠的检测方法。
Type I interferonopathies are characterized by an increase of circulating type I interferon (IFN) concentration. Type I interferonopathies refer to rare Mendelian genetic disorders such as Aicardi-Goutieres Syndrome (AGS) as well as more frequent and polygenic auto-immune diseases like systemic lupus erythematosus (SLE). Yet, detection of type I IFN in these patients remains challenging as its amount is usually very low in patients' sera. Thus, the detection of interferon-stimulating genes has been proposed as an alternative for the detection of this cytokine but sensitivy, specificity and predictive values of the assay have not been reported so far. In this study, we propose two different methods based on Nanostring or RT-qPCR to measure in the clinical routine the IFN response, defined as a set of transcripts that are systemically induced by IFNs. The IFN signature is composed of 6 IFN stimulated genes (ISGs) and has a strong predictive value for the diagnosis of type I interferonopathies. The use of this simple test might represent a gold standard for the evaluation of various autoimmune diseases. Moreover, this test could also be used to monitor patients treated with drugs targeting type I IFN pathway. When comparing both methods - Nanostring and qPCR - in terms of analytical performance, they provided similar results but Nanostring was quicker, easier to multiplex, and almost fully-automated, which represent a more reliable assay for the daily clinical practice.