用BrdU法检测外周血淋巴细胞hprt突变的方法学探讨  

Study on 5-Bromodeoxyuridine Labeling Method for Detecting hprt Mutation in Peripheral Blood Lymphocyte

作　　者：赵涛[1] 崔凤梅[1] 赵经涌[1] 劳勤华[1] 王六一[1] 

机构地区：[1]苏州大学核医学院,江苏苏州215007

出　　处：《工业卫生与职业病》2002年第3期141-144,共4页Industrial Health and Occupational Diseases

基　　金：国防科工委资助项目 (Y5 5 73 0 62 )

摘　　要：目的　探索用 5 -溴脱氧尿嘧啶 (5 bromodeoxyuridine,BrdU)法检测次黄嘌呤鸟嘌呤磷酸核糖转移酶基因 (hypoxanthine guaninephosphoribosytransferasegene,hprt)突变的操作程序。 方法　用正交设计法就BrdU法的最佳检测条件进行了探讨。结果　本研究所得的最佳操作程序是 :将血样放入 4℃冰箱中冷藏2 4h ;培养基中 6 巯基鸟嘌呤 (6 thioguanine ,6 TG)最终浓度为 1×10 -4mol/L ;BrdU最终浓度用1× 10 -4mol/L ;加入BrdU后培养时间为 16h ;Giemsa染液浓度为 4 % ;染色时间为 15min。结论　BrdU法是一种较好的检测hprt突变的方法。Objective 5 Bromodeoxyuridine(BrdU)labeling method is important for detecting hprt mutation.The optimum conditions of this method are experimented.Methods Orthogonal design is used in experiments.Results The optimum procedure is as follows:Keep the blood samples in refrigerator at 4℃ for 24 h .Make the final concentrations of both 6 thioguanine(6 TG)and BrdU in culture medium at 1×10 -4 mol/L .Incubate for 16 h.Stain with 4% Giemsa staining solution for 15 min.Conclusions BrdU labeling method is good for detecting hprt mutation.

关 键 词：BrdU法 检测 外周血 淋巴细胞 hprt突变 正交设计 

分 类 号：R114[医药卫生—卫生毒理学] Q754[医药卫生—公共卫生与预防医学]