cDNA基因芯片技术在血管内皮细胞基因变化研究中的质量保证过程

Analysis of interfered factors about cDNA microarray studying gene expression of human vascular endothelial cells

目的 探讨cDNA基因芯片技术在应用过程中如何排除干扰,保证分析质量,获得进一步归纳分析的有效数据。方法 通过排除非特异本底、点样均一误差、非特异错配、同源组织错配等各方面因素,应用10 368个基因的cDNA基因芯片研究人冠状动脉血管内皮细胞基因表达的总体变化趋势和不同功能集团间的分析,逐步分析不同干扰因素,获得有效处理数据。结果 三次重复试验中10 368个基因芯片反应数据分别排除89、132和116个低于非特异本底荧光强度的杂交点, 281、368和220个少于荧光杂交面积10 %的不确定杂交反应, 1 113、1 026和1 096个不同种属间非特异错配,重复性筛选等因素,获得相对有效的与参考总RNA比较表达高于1 5倍323个,低于1 5倍1 192个,高低1 5倍之间的2 036个数据。结论 基因芯片技术在应用过程中受许多因素的影响,经过切实有效的方法排除干扰因素,可获得较具有客观性的结果,是进一步分析的必要前提。

Objective To study the method for data analysis of gene expression about vascular endothelial cells using cDNA microarray,To prevent from the state or quality of being incapable of producing a desired effect or result. We analyzed the data that cause interference in the special and nonspecial factors.Methods We used a human cardiovascular-based cDNA microarray containing 10 368 genes, Removed the non-special background, spoting error and mis-hybrization between same and different strain.Results In order to detract the background intensity, noise and scale factor , we remove 89 , 132 and 116 spots. 281, 368 and 220 spots that derived from the intensity values of the lowest 10% on the chip. 1 113,1 026 and 1 096 spots from them because mis-blot. Finally we got the 323 spots more than 1.5 times and 1 192 spots lower 1.5 times and 2 036 spots equal to each other.Conclusion Microarray technology has been used as a means of large-scale screening of vast numbers of genes that possess differential expression and interfered by many factors. We must analyzed and removed the interfered factors for example it is important for the availability.