微量全血彗星实验法的建立及在生物监测中的初步应用  

A new approach for directly using micro-whole blood specimen in comet assay and its application in biomonitoring

作　　者：孙树秋[1] 姜树林[1] 姚洪菊[1] 孙殿军[1] 

机构地区：[1]哈尔滨医科大学中国疾病预防控制中心地方病控制中心,黑龙江哈尔滨150086

出　　处：《中国地方病学杂志》2003年第2期170-173,共4页Chinese Jouranl of Endemiology

摘　　要：目的　建立一种简捷、实用的末梢血样直接用于彗星实验的新技术。方法　按随机、对照及重复的实验原则 ,于早、中、晚时段分别采集 6只昆明种雄性小鼠的尾血 ,每次取血样 3 μl/只进行彗星实验。电泳条件设置为电压 2 5V ,电流 13 0mA ,DNA碱性解旋与电泳各 3 0min。并以小样本人群对新建方法的有效性进行了初步验证。结果　①小鼠尾全血彗星图像清晰、对比度高 ,阳性与阴性对照组的拖尾率总体差异明显 ,重现性良好 ;②本方法可用于检测吸烟人群的DNA断裂损伤 ,并能在损伤程度上 (拖尾率与尾长 2项指标明显 )与非吸烟人群相鉴别。结论　基本建立了微量全血彗星实验法 。Objective To establish a simple, rapid and practical method for using micro whole blood specimen in comet assay.Methods Based on the principles of randomization,repeatability and control,6 male mice were tested for establishing a new approach,in which 3 μl tail whole blood from each mouse was collected per time for comet assay at morning,noon and evening,respectively.The electrophoresis conditions were set at 25 V,130 mA,30 min for both DNA unwinding and electrophoresis under pH13.And the newly developed method was used in a small size human population for validation.Results ①Comet image is clear and high contrast.And the percentage of tailed cells was notably different between positive group and negative group; ②the newly developed method was preliminarily proved to be valid in distinguishing smokers and nonsmokers,particularly in such indicators as percentage of tailed cells,and DNA tail length of comet cells.Conclusions The new approach for using micro whole blood specimen in comet assay has been preliminarily established in our laboratory.It is,however,need being modified for more practical use in biomonitoring.

关 键 词：微量全血 实验法 初步应用 生物监测 DNA断裂损伤 彗星实验 非吸烟人群 雄性小鼠 条件设置 图像清晰 损伤程度 取血样 昆明种 小样本 对照组 对比度 重现性 实用性 电泳 验证 拖尾 

分 类 号：R115[医药卫生—公共卫生与预防医学]