An evaluation of the performance of cDNA microarrays for detecting changes in global mRNA expression
An evaluation of the performance of cDNA microarrays for detecting changes in global mRNA expression
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DOI：
10.1093/nar/29.8.e41
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发表时间：
2001-04-15
影响因子：
14.9
通讯作者：
Johnston, Rick
中科院分区：
文献类型：
--
作者：
Yue, Huibin;Eastman, P. Scott;Johnston, Rick
cDNA微阵列是一种有潜力精确测量全球mRNA表达水平变化的技术方法。我们报告了一个优化的cDNA微阵列平台的评估，以产生准确，精确和可靠的数据与使用微阵列作为采集平台，以填充基因表达数据库的目标一致。该研究设计包括两项独立评价，使用来自两个不同生产批次的70个阵列，并使用三种人体组织来源作为样本：胎盘、大脑和心脏。总的信号响应在三个数量级上是线性的，并且对任何元素的灵敏度估计为2 pg mRNA。所有未分化元件的差异表达的计算变异系数在整个信号范围内为12-14%，并且不随阵列批次或组织来源而变化。差异表达的最小可检测倍数变化为1.4。就偏倚（观察值减去预期差异表达比率）而言，所有未分化元素的准确度均小于1/10000。本报告中呈现的结果证明了cDNA微阵列技术平台的可再现性能，并且该方法为评估监测全球mRNA表达变化的其他技术提供了有用的框架。
The cDNA microarray is one technological approach that has the potential to accurately measure changes in global mRNA expression levels. We report an assessment of an optimized cDNA microarray platform to generate accurate, precise and reliable data consistent with the objective of using microarrays as an acquisition platform to populate gene expression databases. The study design consisted of two independent evaluations with 70 arrays from two different manufactured lots and used three human tissue sources as samples: placenta, brain and heart. Overall signal response was linear over three orders of magnitude and the sensitivity for any element was estimated to be 2 pg mRNA. The calculated coefficient of variation for differential expression for all non-differentiated elements was 12-14% across the entire signal range and did not vary with array batch or tissue source. The minimum detectable fold change for differential expression was 1.4. Accuracy, in terms of bias (observed minus expected differential expression ratio), was less than 1 part in 10 000 for all non-differentiated elements. The results presented in this report demonstrate the reproducible performance of the cDNA microarray technology platform and the methods provide a useful framework for evaluating other technologies that monitor changes in global mRNA expression.