毛细管电泳基因扫描和凝胶电泳异源双链分析在免疫球蛋白/T细胞受体基因重排检测中的应用研究  

Comparative study of capillary electrophoresis-genescan and polyacrylamide gel electrophoresis-heteroduplex polymorphism in detecting IG/TCR gene rearrangement

作　　者：陈杰[1] 郑可[1] 张文燕[1] 孙林雍 唐源[1] 严嘉琦 赵莎[1] 刘卫平[1] 

机构地区：[1]四川大学华西医院病理科,四川成都610041

出　　处：《西部医学》2016年第11期1489-1494,共6页Medical Journal of West China

基　　金：国家自然科学基金(81272626)

摘　　要：目的探讨免疫球蛋白轻链IgK及T细胞抗原受体（TCRT）基因重排更有效的检测方法。方法收集四川大学华西医院病理科2013～2014年淋巴组织增生性病变石蜡样本共139例，采用B10MED2系统扩增，分别使用毛细管电泳基因扫描和凝胶电泳异源双链分析两种方法进行IgK和TCR7基因重排检测，比较不同方法之间的检出率。结果81例行IgK基因重排检测，其中59例病理诊断为B细胞淋巴瘤，毛细管电泳基因扫描检出阳性47例（79．66％），凝胶电泳异源双链分析检出阳性34例（57．63％），二者差异有统计学意义（P=0．01）。58例行TCR7基因重排检测，其中26例病理诊断为T细胞淋巴瘤，毛细管电泳基因扫描检出阳性21例（80．77％），凝胶电泳异源双链检出阳性17例（65．38％），差异无统计学意义（P=0．211）。结论毛细管电泳基因扫描在抗原受体基因重排检测中比凝胶电泳异源双链分析有更好的检测灵敏度。Objective To compare the sensitivity of immunoglobulin heavy chain （IGH） gene rearrangement, immunoglohulin light chain（IgK） rearrangement and TCRy gene clonal rearrangment by capillary eleetrophoresis （CE）- genescan and polyacrylamide gel electrophoresis （PAGE）-heteroduplex polymorphism, then establish a more effective method in detecting gene rearrangement. Methods DNA was extracted from paraffin embedded tissues in 139 patients of lymphoproliferative disease which collected from Department of Pathology, West China Hospital of Sichuan University in the period of 2013 to 2014. Clonal gene rearrangement was detected by PAGE-heteroduplex poiymorphism and CE-genes- can and compare the sensitivity of the two methods. Results 81 patients were positive for IgK rearrangement. 59 cases were diagnosed as B-cell lymphomas. 47 cases （79.66%） were positive for IgK rearragement by CE-genescan, whereas the positive rate was 57.63% by PAGE-heteroduplex polymorphism. 58 cases were positive for T-cell receptor （TCRy） gene rearrangement. 26 of these were diagnosed as T-cell lymphomas. 80.77% （21/26） and 65.38% （17/26） TCRy clonality were detected by CE-genescan and by PAGE-heteroduplex polymorphism, respectively. In composition, The positive rate of the former was higher than that of the latter, and there was a significant difference in the IgH rearrangement （P=0.01）, while there was no significant difference in TCRy rearrangement （P= 0. 211 ）. Conclusion CE-genescan has a higher detecting sensitivity to antigen receptor gene rearrangement than PAGE-heteroduplex polymorphism. It＇s an efficient detection of gene rearrangement.

关 键 词：毛细管电泳 凝胶电泳 基因重排 BIOMED-2系统 

分 类 号：R-331[医药卫生] R733