Simple and precise detection of UGT1A1 polymorphisms with a modified loop-hybrid mobility shift assay using Cy5-labeled loop probes
Simple and precise detection of UGT1A1 polymorphisms with a modified loop-hybrid mobility shift assay using Cy5-labeled loop probes
复制标题
使用 Cy5 标记的环探针，通过改良的环混合迁移率变动分析简单而精确地检测 UGT1A1 多态性
DOI：
10.1016/j.cca.2011.05.021
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
and Miyagi Y
中科院分区：
文献类型：
--
作者：
Matsukuma S;Saito H;Yamada K;Okamoto N;Ishikawa T;Yoshihara M;Koizume S;Sakuma Y;and Miyagi Y
吡立替康是一种拓扑异构酶I抑制剂，作为重要的抗癌治疗药物已被广泛应用。已知该药对过敏患者的有害作用与ugt1a1基因的遗传多态性有关，即多态变体，28和6。方法采用cy5标记的环杂化探针，提出了一种改进形式的环杂化迁移转移测定法，作为一种精确、简便的方法，测定了在28位点的TA重复多态性。结果在改进的方法中，cy5荧光信号只检测到环杂交条带，尽管由于PCR产物中TA重复而出现了不规则的电泳条带。当将使用cy5标记探针的环杂交组合在一起并用于迁移位移测定时，可以在单车道内实现对<s:2> / 28和<s:2> / 6变体的同时分型。
BackgroundIrinotecan, an inhibitor of topoisomerase I, has been widely used as an important anti-cancer therapeutic drug. Deleterious effects of the drug in hypersensitive patients are known to be associated with genetic polymorphisms of theUGT1A1gene, namely the polymorphic variants,⁎28 and⁎6.MethodsA modified form of loop-hybrid mobility shift assay using a Cy5-tagged loop-hybrid probe was proposed as a precise and easy method of determining TA repeat polymorphisms at the⁎28 locus.ResultsIn this modified method, only loop-hybrid bands were detected by a Cy5-fluorescent signal, despite several irregular electrophoretic bands due to TA repeats in the PCR product.ConclusionsWhen a loop-hybrid using a Cy5-tagged probe for the⁎28 locus and⁎6 locus were combined and used for mobility shift assay, simultaneous typing of the⁎28 and⁎6 variants was achieved in a single lane.