PMC:3091642 / 12749-14095 JSONTXT

{"project":"2_test","denotations":[{"id":"20663125-11846609-10592586","span":{"begin":309,"end":311},"obj":"11846609"}],"text":"To validate the microarray data, 20 genes were subjected to qRT-PCR using gene specific primer pairs and the same RNA samples as in the microarray analysis (Table 2). Results were analyzed by 2-∆∆Ct method to determine relative levels of gene expression at each dpi time point compared to uninfected control [31]. There were no differences between microarray data and the qRT-PCR at any dpi time point (Table 3). However, it should be noted that fold change values for certain genes obtained by qRT-PCR analysis showed much greater expression levels than those observed in the microarray analysis. For example, the fold changes for the gene expression of matrix metalloproteinase (MMP) 27, interleukin (IL) 6, fatty acid binding protein (FABP) 4, IL8, and CXC chemokine K60 at 3- or 5 dpi showed much higher levels in qRT-PCR analysis compared to fold changes shown in microarray analysis (Table 3). Possibly, this qualitative difference between methodologies may be attributed to the upper detection limits of the fluorescent intensities for the array scanner. Based on quality control measures, such as the spike-in controls and the results of targeted qRT-PCR indicate that the microarray data sets for differential gene expression are valid to investigate genome-wide differential expression patterns for host responses during ILTV infection."}