Biomarkers for monitoring pre-analytical quality variation of mRNA in blood samples.
Biomarkers for monitoring pre-analytical quality variation of mRNA in blood samples.
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DOI：
10.1371/journal.pone.0111644
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Rian E
中科院分区：
文献类型：
--
作者：
Zhang H;Korenková V;Sjöback R;Švec D;Björkman J;Kruhøffer M;Verderio P;Pizzamiglio S;Ciniselli CM;Wyrich R;Oelmueller U;Kubista M;Lindahl T;Lönneborg A;Rian E
在分子诊断工作流程的分析前阶段，越来越需要适当的质量控制工具。本研究的目的是确定两种不同类型的采血管，K2EDTA （EDTA）管和PAXgene blood RNA管（PAXgene管）中监测分析前mRNA质量变化的生物标志物。这些试管广泛用于诊断设置以及研究生物库样本。在两种不同的采血管中采集的血液标本在不同的温度下保存不同的时间，并对所得提取的RNA进行微阵列分析。我们发现了大量潜在的mRNA质量生物标志物，可用于监测采血后基因表达变化和血液中mRNA的降解。生成潜在生物标记物和一组相关内参基因的qPCR分析，并用于预先验证所选生物标记物的子集。确定了潜在的qPCR生物标志物的检测精度，并使用所开发的qPCR检测方法和来自60名健康受试者的血液样本对所选择的优质生物标志物进行了最终验证。总共成功验证了4个mRNA质量生物标志物（USP32、LMNA、FOSB、TNRFSF10C）。我们建议使用这些血液mRNA质量生物标志物来验证实验前分析工作流程。这些生物标记物在SPIDIA-RNA项目的第二环试验中被进一步评估，证明这些生物标记物可以用作血液样本mRNA分析的质量控制工具。
There is an increasing need for proper quality control tools in the pre-analytical phase of the molecular diagnostic workflow. The aim of the present study was to identify biomarkers for monitoring pre-analytical mRNA quality variations in two different types of blood collection tubes, K2EDTA (EDTA) tubes and PAXgene Blood RNA Tubes (PAXgene tubes). These tubes are extensively used both in the diagnostic setting as well as for research biobank samples. Blood specimens collected in the two different blood collection tubes were stored for varying times at different temperatures, and microarray analysis was performed on resultant extracted RNA. A large set of potential mRNA quality biomarkers for monitoring post-phlebotomy gene expression changes and mRNA degradation in blood was identified. qPCR assays for the potential biomarkers and a set of relevant reference genes were generated and used to pre-validate a sub-set of the selected biomarkers. The assay precision of the potential qPCR based biomarkers was determined, and a final validation of the selected quality biomarkers using the developed qPCR assays and blood samples from 60 healthy additional subjects was performed. In total, four mRNA quality biomarkers (USP32, LMNA, FOSB, TNRFSF10C) were successfully validated. We suggest here the use of these blood mRNA quality biomarkers for validating an experimental pre-analytical workflow. These biomarkers were further evaluated in the 2nd ring trial of the SPIDIA-RNA Program which demonstrated that these biomarkers can be used as quality control tools for mRNA analyses from blood samples.
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影响因子：
2.7
作者：
Opitz L;Salinas-Riester G;Grade M;Jung K;Jo P;Emons G;Ghadimi BM;Beissbarth T;Gaedcke J
通讯作者：
Gaedcke J
影响因子：
2.2
作者：
Bauer, M;Polzin, S;Patzelt, D
通讯作者：
Patzelt, D
DOI：
10.1007/978-1-60327-353-4_1
发表时间：
2013-01-01
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Johnson, Gemma;Nolan, Tania;Bustin, Stephen A
通讯作者：
Bustin, Stephen A
影响因子：
5.7
作者：
Funk, Dorothy M. Adcock;Lippi, Giuseppe;Favaloro, Emmanuel J.
通讯作者：
Favaloro, Emmanuel J.
影响因子：
4.1
作者：
Betsou, Fotini;Gunter, Elaine;Chuaqui, Rodrigo
通讯作者：
Chuaqui, Rodrigo