固定剂及固定时间对DNA提取质量的影响  

Effect of different fixatives and fixation tune on DNA quality and quantity

作　　者：李海军[1] 柳刚[1] 蒋会勇[1] 贺莉[1] 彭宏[1] 朱梅刚[1] 赵彤[1] 

机构地区：[1]第一军医大学南方医院病理科,广州510515

出　　处：《诊断病理学杂志》2004年第3期182-184,i013,共4页Chinese Journal of Diagnostic Pathology

基　　金：广东省科技计划项目(B30101);广州科技计划项目(2002Z3-E4061)

摘　　要：目的 探讨10％福尔马林、95％乙醇、中性缓冲福尔马林、4％多聚甲醛-0.1 moL/L磷酸缓冲液4种固定剂对新鲜组织DNA质量及PCR的影响。方法 选取16例单纯性扁桃体炎组织标本的冷冻切片,进行不同时段的固定,比较提取物OD260nm/OD280nm比值、DNA浓度和PCR结果。结果 各组之间同一时间、同一组内不同时间OD260nm/OD280nm比值比较差异不显著(P>0.05);而同一固定时间段,个别组间DNA浓度以及同一组内不同时间DNA浓度差异显著(P<0.05)。固定72h后,各组的PCR阳性率分别为66.7％、100％、93.3％、80％、100％。结论95％乙醇固定效果最佳,其余3组固定剂随固定时间的延长,PCR阳性率逐渐降低。Objective To investigate the effects of four different fixatives on the quality and quantity of DNA and the subsequences of PCR. Methods 16 samples obtained from simple tonsillitis fresh tissue were fixed for five different hours. The ratio of OD260nm/OD280nm, concerntration of DNA and results of PCR were compared. Results Among each extractive, no significant differences was delected in OD260nm/OD280nm(P > 0.05) , but a significant difference was found in content of DNA( P < 0.05) . In 72 h period, (he positive ratio were 66.7% , 100% , 93.3% , 80 % and 100 % . Conclusion 95% ethanol is the best fixative in the amplification of an objective fragment in fresh lymphoid tissue. With the increasing of fixation time, the positive ratio decrease in the other three fixatives.

关 键 词：固定剂 固定时间 DNA 提取质量 影响 淋巴组织 淋巴瘤 病理诊断 

分 类 号：R733.4[医药卫生—肿瘤]