PCR技术在检测sp3基因表达和ApoE基因型中的应用  

Applying PCR Technique to Examine the Gene Expression of sp3 and ApoE Genotype

作　　者：乔立艳[1] 许贤豪[1] 冯颖[1] 张华[1] 殷剑[1] 耿莹[1] 魏雪飞[1] 王红[1] 国红[1] 侯世芳[1] 文诗广[1] 刘江红[1] 矫毓娟[1] 

机构地区：[1]卫生部北京医院神经内科,北京100730

出　　处：《中国神经免疫学和神经病学杂志》2003年第1期20-24,共5页Chinese Journal of Neuroimmunology and Neurology

基　　金：国家自然科学基金资助项目 (3 9870 2 77)

摘　　要：目的　应用多聚酶链式反应 (PCR)技术检测多发性硬化 (MS)和老年性痴呆 (AD)的相关基因 sp3和Apo E。方法　分离 MS患者外周血单个核细胞 (PBMC) ,提取 RNA,反转录合成 c DNA第 1链 ,PCR扩增 sp3基因片段 ,应用反转录 -PCR(RT-PCR)技术在 RNA水平上检测 sp3基因在 MS患者 PBMC的表达情况 ;取 AD患者外周血 ,用低渗溶血法提取白细胞 ,酚 /氯仿法提取 DNA,应用 PCR-限制性片段长度多态性 (PCR-RFLP)分析技术检测 Apo Eε基因的多态性。结果　MS组 sp3阴性表达率 (4 5 % )明显高于对照组 (P <0 .0 1 )。AD患者的 Apo Eε2、ε3、ε4基因频率依次为 0 .0 67、 0 .70 0、 0 .2 3 3 ,AD患者的 Apo Eε4基因频率明显高于正常人。结论　汉族人 MS患者存在 sp3基因表达缺陷 ;Apo Eε4与 AD密切相关 ;PCR相关技术可作为基因研究的有力工具。Objective Polymerase chain reaction (PCR ) technique was used to test sp3 and Apolipoprotein E(ApoE) genes which correlate with multiple sclerosis(MS) and Alzheimer disease (AD). Methods Isolation of the peripheral mononuclear cells. Extraction of RNA and synthesized the first strand of cDNA,to amplify the sp3 fragments with PCR technique for examining the expression status of sp3 in MS patients. Investigated ApoE genotypes with PCR and restriction fragment length polymorphism (RFLP) with extracted DNA from the peripheral blood with the phenol chloroform procedure in AD patients. Results The frequency of deficient expression of sp3 in MS patients is significantly higher than that in control groups (P<0.01). The frequencies of ApoE ε2 ,ε3 and ε4 were 0 067 ,0 700 and 0 233 respectively in AD. The frequency of ε4 was significantlyhigher in AD than that in controls. Conclusions Our results indicate that sp3 and ApoE ε4 are correlated with development of MS and AD, respectively. PCR is useful as a powerful tool for genetic study.

关 键 词：SP3 APOE RT-PCR PCR-RFLP 多发性硬化 阿尔茨海默病 

分 类 号：R446[医药卫生—诊断学]