Dallas Peter B, Gottardo Nicholas G, Firth Martin J, Beesley Alex H, Hoffmann Katrin, Terry Philippa A, Freitas Joseph R, Boag Joanne M, Cummings Aaron J, Kees Ursula R
Division of Children's Leukaemia and Cancer Research, Telethon Institute for Child Health Research and Centre for Child Health Research, The University of Western Australia, Perth, Australia.
BMC Genomics. 2005 Apr 27;6:59. doi: 10.1186/1471-2164-6-59.
The use of microarray technology to assess gene expression levels is now widespread in biology. The validation of microarray results using independent mRNA quantitation techniques remains a desirable element of any microarray experiment. To facilitate the comparison of microarray expression data between laboratories it is essential that validation methodologies be critically examined. We have assessed the correlation between expression scores obtained for 48 human genes using oligonucleotide microarrays and the expression levels for the same genes measured by quantitative real-time RT-PCR (qRT-PCR).
Correlations with qRT-PCR data were obtained using microarray data that were processed using robust multi-array analysis (RMA) and the MAS 5.0 algorithm. Our results indicate that when identical transcripts are targeted by the two methods, correlations between qRT-PCR and microarray data are generally strong (r = 0.89). However, we observed poor correlations between qRT-PCR and RMA or MAS 5.0 normalized microarray data for 13% or 16% of genes, respectively.
These results highlight the complementarity of oligonucleotide microarray and qRT-PCR technologies for validation of gene expression measurements, while emphasizing the continuing requirement for caution in interpreting gene expression data.
利用微阵列技术评估基因表达水平如今在生物学领域已广泛应用。使用独立的mRNA定量技术对微阵列结果进行验证仍是任何微阵列实验中理想的环节。为便于各实验室之间比较微阵列表达数据，对验证方法进行严格审查至关重要。我们评估了使用寡核苷酸微阵列获得的48个人类基因的表达分数与通过定量实时RT-PCR（qRT-PCR）测量的相同基因表达水平之间的相关性。
使用经过稳健多阵列分析（RMA）和MAS 5.0算法处理的微阵列数据获得了与qRT-PCR数据的相关性。我们的结果表明，当两种方法针对相同转录本时，qRT-PCR与微阵列数据之间的相关性通常很强（r = 0.89）。然而，我们分别观察到，对于13%或16%的基因，qRT-PCR与RMA或MAS 5.0标准化微阵列数据之间的相关性较差。
这些结果凸显了寡核苷酸微阵列和qRT-PCR技术在验证基因表达测量方面的互补性，同时强调在解释基因表达数据时仍需持续谨慎。
