Multicenter evaluation of a standardized protocol for noninvasive gene expression profiling.
Multicenter evaluation of a standardized protocol for noninvasive gene expression profiling.
复制标题
DOI：
10.1111/ajt.12284
复制
发表时间：
2013-07
期刊：
影响因子：
--
通讯作者：
Fairchild RL
中科院分区：
文献类型：
--
作者：
Keslar KS;Lin M;Zmijewska AA;Sigdel TK;Tran TQ;Ma L;Bhasin M;Rao P;Ding R;Iklé DN;Mannon RB;Sarwal MM;Strom TB;Reed EF;Heeger PS;Suthanthiran M;Fairchild RL
移植受体血液和尿液的基因表达谱可以潜在地用于监测移植物功能，但使用的众多方案使得共享数据和比较来自不同实验室的结果变得困难。本研究的目的是评估当前RNA分离、逆转录和定量聚合酶链反应（qPCR）方法的性能，并测试多个中心使用标准化方案是否可以获得相同的结果。将样品、试剂和详细说明分发到6个参与位点，按指示对18S、PRF、GZB、IL8、CXCL9和CXCL10进行RNA分离、反转录和qPCR。所有数据均在同一地点进行分析。所有位点均表现出RNA分离和qPCR分析的熟练程度。所有靶基因表达量与样品的相关系数为>0.938。样本对间折叠变化的变异系数小于40%。所有站点都能够在1.5的因子范围内准确地量化已知浓度的对照样本。总的来说，我们已经制定并验证了测量血液和尿液中基因表达的详细方法，这些方法可以在多个实验室中产生一致的结果。
Gene expression profiling of transplant recipient blood and urine can potentially be used to monitor graft function, but the multitude of protocols in use make sharing data and comparing results from different laboratories difficult. The goal of this study was to evaluate the performance of current methods of RNA isolation, reverse transcription, and quantitative polymerase chain reaction (qPCR) and to test whether multiple centers using a standardized protocol can obtain the same results. Samples, reagents, and detailed instructions were distributed to six participating sites that performed RNA isolation, reverse transcription and qPCR for 18S, PRF, GZB, IL8, CXCL9 and CXCL10 as instructed. All data were analyzed at a single site. All sites demonstrated proficiency in RNA isolation and qPCR analysis. Gene expression measurements for all targets and samples had correlations >0.938. The coefficient of variation of fold-changes between pairs of samples was less than 40%. All sites were able to accurately quantify a control sample of known concentration within a factor of 1.5. Collectively, we have formulated and validated detailed methods for measuring gene expression in blood and urine that can yield consistent results in multiple laboratories.
登录
查看更多内容
影响因子：
19.6
作者：
Aquino-Dias, E. C.;Joelsons, G.;Manfro, R. C.
通讯作者：
Manfro, R. C.
影响因子：
6.2
作者：
Dadhania, D;Muthukumar, T;Suthanthiran, M
通讯作者：
Suthanthiran, M
影响因子：
5.7
作者：
Mas, Valeria R.;Mas, Luciana A.;Maluf, Daniel G.
通讯作者：
Maluf, Daniel G.
影响因子：
3.7
作者：
Ozbay, A.;Torring, C.;Carstens, J.
通讯作者：
Carstens, J.
影响因子：
6.2
作者：
Sabek, M;Dorak, MT;Gaber, L
通讯作者：
Gaber, L