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目的 用免疫相关基因芯片寻找多发性硬化表达差异的免疫相关基因。方法 将 484条免疫相关基因的double(双点 )cDNA产物按微矩阵排列点样于玻片上。例 1患者及其对照为第 1组 ,例 2患者及其对照为第 2组 ,例 3患者及其对照为第 3组。取各组外周血 ,分离淋巴细胞 ,提取总RNA ,用RT PCR逆转录成cDNA。标记cDNA探针 ,分别用cy3 dUTP、cy5 dUTP标记正常人和多发性硬化患者cDNA。将基因芯片和杂交探针变性后杂交、洗干。用ScanArray 40 0 0扫描芯片 ,GenePixPro3 0软件分析cy3、cy5两种荧光信号的强度和比值。结果 1组中筛选出差异表达的基因 2 2项 ,2组中出现差异表达的基因 2 7项 ,3组中出现差异表达的基因 72项。具有同一GeneID号的一对Double基因 ,其中 1组 6对 ,2组 9对 ,3组 30对。结论 试验结果显示正常人与MS患者免疫相关基因的表达有显著差异 ;不同发病阶段免疫相关基因表达有差异 ;以细胞免疫相关基因变化为主 ,体液免疫相关基因亦有异常表达
Objective To search for immune related genes with different expression of multiple sclerosis by immune related gene chip. Methods The double (double-dot) cDNA products of 484 immune-related genes were microarrayed on glass slides. Example 1 Patients and their controls were Group 1, patients in Group 2 and their controls were Group 2, patients in Group 3 and Group 3 were controls. Peripheral blood was taken from each group, lymphocytes were isolated, total RNA was extracted and reverse transcribed into cDNA by RT PCR. The cDNA probes were labeled and cy3 dUTP and cy5 dUTP were used to label normal and multiple sclerosis patients. The gene chip and hybridization probe denatured hybridization, washed and dried. With ScanArray 40 0 0 scanning chip, GenePixPro30 software analysis cy3, cy5 two kinds of fluorescence signal intensity and ratio. Results A total of 22 differentially expressed genes were screened in group 1, 27 differentially expressed genes in 2 groups and 72 differentially expressed genes in 3 groups. A pair of Double Genes with the same GeneID, including 6 pairs in 1 set, 9 pairs in 2 sets and 30 pairs in 3 sets. Conclusions The results showed that there was a significant difference in the expression of immune-related genes between normal subjects and MS patients. The expression of immune-related genes was different at different stages of disease. The changes of cellular immunity-related genes were also predominant, and the genes related to humoral immunity were also abnormally expressed