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通过PAXgene和Tempus全血RNA采集与分离系统进行的I型干扰素评分测定比较。

Comparable type I interferon score determination from PAXgene and Tempus whole blood RNA collection and isolation systems.

作者信息

Lamot Lovro, Niemietz Iwona, Brown Kelly L

机构信息

Division of Rheumatology, Department of Pediatrics, Faculty of Medicine, The University of British Columbia, Vancouver, BC, Canada.

BC Children's Hospital Research Institute, Rm A4-145, 950 West 28th Ave, Vancouver, BC, V5Z 4H4, Canada.

出版信息

BMC Res Notes. 2019 Aug 15;12(1):511. doi: 10.1186/s13104-019-4562-z.

Abstract

OBJECTIVE

Type I interferons (IFN) have important roles in many immune-mediated inflammatory diseases (IMIDs) and are a relatively new therapeutic target. Direct detection of type I IFNs has proved challenging, thus their presence is often inferred from the expression of interferon-stimulated genes (ISGs) and calculation of an interferon score (IS). The objective of this research was to determine if the expression of six common ISGs and subsequent IS were comparable when RNA was derived from the Tempus and PAXgene whole blood RNA collection systems.

RESULTS

Whole blood was obtained from ten healthy adults, incubated ex vivo in the absence and presence of recombinant human IFNα then divided into PAXgene and Tempus tubes. Despite reports of tube-specific patterns of gene expression, quantitative PCR (qPCR) analysis revealed no significant differences between PAXgene and Tempus tubes in either the homeostatic or IFNα-induced expression of six ISGs (IFI27, IFI44L, IFIT1, ISG15, RSAD2, SIGLEC1). Overall there was a strong correlation in the IS between unstimulated (r = 0.92, p = 0.0005) and IFNα-stimulated (r = 0.71, p = 0.0268) samples derived from the PAXgene and Tempus tubes.

摘要

目的

I型干扰素(IFN)在许多免疫介导的炎症性疾病(IMID)中发挥重要作用,是一个相对较新的治疗靶点。直接检测I型干扰素已被证明具有挑战性,因此其存在通常通过干扰素刺激基因(ISG)的表达和干扰素评分(IS)的计算来推断。本研究的目的是确定当RNA来自Tempus和PAXgene全血RNA收集系统时,六种常见ISG的表达及随后的IS是否具有可比性。

结果

从十名健康成年人采集全血,在体外分别于不存在和存在重组人IFNα的情况下孵育,然后分为PAXgene管和Tempus管。尽管有关于特定管型基因表达模式的报道,但定量PCR(qPCR)分析显示,在六种ISG(IFI27、IFI44L、IFIT1、ISG15、RSAD2、SIGLEC1)的稳态或IFNα诱导表达方面,PAXgene管和Tempus管之间没有显著差异。总体而言,来自PAXgene管和Tempus管的未刺激样本(r = 0.92,p = 0.0005)和IFNα刺激样本(r = 0.71,p = 0.0268)的IS之间存在很强的相关性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cd2/6694656/de64f21ec4aa/13104_2019_4562_Fig1_HTML.jpg

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