冻存血与新鲜血的DNA抽提效果的比较研究

Comparative study of the effect on extracted DNA between fresh and frozen whole blood

目的探讨一种简单便捷、可用于PCR扩增的人体外周血标本冷冻保存的方法。方法外周血标本冷藏于普通冰箱冷冻室2-4周,采用常规方法,提取28例新鲜血标本及31例冻存血标本的DNA,经2%琼脂糖凝胶电泳,凝胶分析软件半定量分析,比较冻存血和新鲜血的DNA抽提效果。采用PCR方法扩增抽提DNA标本的ABCB4基因外显子6,研究冻存血中提取的DNA能否用于PCR实验。结果新鲜血组的光强度值为24948.98±10246.97,冻存血组DNA标本的光强度值为22917.65±11545.63,两组间无显著性差异(P>0.05)。所有冻存血中提取的DNA标本,经PCR扩增均有目标带。结论简易冷冻保存的外周血标本中提取的DNA质量与新鲜血无明显差别。冻存血的DNA标本可以用于PCR实验。

Objective In an attempt to explore a simple and convenient approach to store blood sample with frozen pack storage that can be used in polymerase chain reaction (PCR) amplification. Methods The specimens of whole blood were reserved in refrigerating chamber of domestic refrigerator in 2 to 4 weeks followed DNA extracting from 28 fresh and 31 frozen specimens by routine method. Subsequently 2% agarose gel elec trophoresis technique and the software of quantitative analysis were employed to compare the advantage of extracted DNA between fresh and frozen blood. The extracted DNA specimens from frozen blood were amplified for exon 6 of ABCB4 gene by PCR to evaluate whether it could be used in the PCR amplification. Results The value of intensity were 24948.98 ± 10246. 97 in fresh blood group, and 22917. 65±11545. 63 in frozen blood group. There were no significant difference between two groups(P>0.05). All extracted DNA specimens from frozen blood visualized the aim band by PCR. Conclusion The findings in extracted DNA reveal no significant difference between fresh blood and plain frozen pack storage, which suggests that DNA from frozen blood can be used in PCR.