标本因素对全自动磁珠法全血DNA提取的影响  

The quality of automatic extracted DNA associated with blood sample status

作　　者：李王霞[1] 刘光箭[1] 陆华新[1] 沈钢[1] 朱远雁[1] 邹洁[1] 

机构地区：[1]武汉血液中心,湖北武汉430030

出　　处：《中国输血杂志》2014年第1期29-31,共3页Chinese Journal of Blood Transfusion

摘　　要：目的探讨抗凝是否完全、冷藏时间长短及冻融次数这3种标本因素对全自动磁珠法提取全血DNA浓度和纯度的影响。方法全血以EDTA抗凝。290份新鲜冷冻标本按抗凝是否完全分完全抗凝组(231份)和不完全抗凝组(59份)。330份完全抗凝标本按冷冻时间分新鲜冷冻组(-20℃冰冻<7 d,231份)、新鲜冷藏组(4℃冷藏<7 d,62份)与陈旧冷藏组(4℃冷藏>24 d,37份)。32份完全抗凝新鲜冷冻标本反复冻融后提取DNA,按冻融次数分1、2、3次冻融。采用全自动磁珠法提取全血DNA,利用DNA定量仪测定DNA浓度与纯度(OD260/280)。统计分析各组DNA浓度与纯度间差异。结果完全抗凝组DNA合格率(98.7%),不完全抗凝组(66.1%);新鲜冷冻组、新鲜冷藏组与陈旧冷藏组DNA浓度、纯度呈递减趋势;1次冻融DNA浓度和纯度高于2次冻融和3次冻融;2次冻融DNA纯度高于3次冻融,但浓度下降不显著。结论抗凝不完全、冷藏时间长及冻融次数多均会使DNA量和纯度下降。相比较而言,抗凝不完全和冷藏时间长影响更大。Objective To explore the related factors for a better DNA extraction,such as anticoagulation status,time of cold storage, and refreezing times before extracting DNA from wl^ole blood. Methods All whole blood specimens were anti- coagulate by EDTA. And all specimens were grouped by three factors mentioned above. The groups were classified as fully anticoagulating group （ n = 231 ）, half anticoagulating group （ n = 59 ）, fresh cold storage group （ n = 231, - 20~C, within 7 days ）, fresh refrigerated group （ n = 62,4~C, within 7 days）, obsolete refrigerated group （ n = 62, 4~C, more than 24 days ） , and repetitive freeze-thawing group （ n = 32）. DNA was extract by automated instrument base on magnetic beads method. DNA concentration and purity were detected, and then submitted to statistical analysis. Results The DNA Concentration and purity of full anticoagulating group is higher than half anticoagulating groups. Meanwhile, the DNA concentration and purity is also going down along with fresh refrigerated group,fresh frozen group,and obsolete refrigerated group. Except for no significant decline between two and three times freeze-thawing, the DNA concentration and purity is going down when samples refreeze more times. Conclusion Half anticoagulating,long refrigerating time and too much freeze-thawing all affect DNA concentration and purity,while anticoagulation and refrigerating time contribute the most.

关 键 词：DNA 标本 抗凝 冷藏 冻融 

分 类 号：E457.11[军事—军事理论]