改进的反向斑点杂交方法  

Improved method in reverse dot blot hybridization

作　　者：林炳生[1,2] 张太松[2] 胥顺[2] 董瑞华[1,2] 陈华云[2] 李明 程钢[1,2] 

机构地区：[1]中山大学药学院,广东广州510006 [2]中山大学达安基因诊断中心,广东广州510665

出　　处：《分子诊断与治疗杂志》2011年第1期18-21,共4页Journal of Molecular Diagnostics and Therapy

基　　金：国家科技重大专项(2008ZX10001-013)

摘　　要：目的改进反向斑点杂交的操作方法,探索更为简便快捷的实验流程。方法优化杂交液II的组分,使用HPV基因分型、β-地中海贫血基因突变和HBV基因分型三个试剂盒验证改进的实验方法。结果 HPV基因分型、β-地中海贫血基因突变和HBV基因分型试剂盒的实验结果证实,使用优化后的杂交液II,链霉亲和素过氧化物酶在杂交温度能够有效地与生物素结合,试剂盒的灵敏度不受影响且膜条的背景低。结论改进的反向斑点杂交方法不影响试剂盒的灵敏度,并且操作简便、实验时间短、实验结果背景低。Objective To improve the operation method of reverse dot blot hybridization as well as explore a simple and quick experimental process. Methods The hybridzation solution components were optimized, and use three kits ofHPV genotyping, β-thalassemia gene mutation and HBV genotyping to validate the improved method. Results Three kits ofHPV genotyping, β-thalassemia gene mutation and HBV genotyping all verify that Streptavidin-POD in the optimized hybrid fluid components can bind biotin effectively at hybridization temperature, and the result shows that the sensitivity of kits is not affected and the background of membrane is lower. Conclusion Improved method of reverse dot blot hybridization does not affect the sensitivity of kits. The operation is easier and eosts less time as well as the background of experiment results is lower.

关 键 词：反向斑点杂交 链霉亲和素过氧化物酶 操作简便 

分 类 号：R440[医药卫生—诊断学]