不同保存时间对于血液DNA提取质量的影响
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摘要
目的探讨不同保存时间对于血液DNA提取质量的影响。方法将新鲜抽取的血液离心得上清,分别冻存1年、2年、3年和4年;之后进行DNA提取,并对提取的DNA的完整性和纯度等进行分析。结果冻存1年的白膜样本与新鲜全血白膜相比,其得率下降极显著,差异具有统计学意义(P <0.01);冻存2年的白膜样本与冻存1年的白膜样本相比,其得率下降呈显著水平,差异具有统计学意义(P<0.05);冻存2,3,4年的白膜样本得率比较,差异无统计学意义(P> 0.05)。冻存时间对于样本的提取纯度无显著性影响;冻存时间对于DNA的完整性无影响。结论在-80℃保存的白膜样本,在最初冻存时有可能会造成DNA数量上的损失,但对于纯度及完整性影响不大。
Objective To investigate the effect of blood retention time on the quality of DNA extraction of blood. Methods The supernatant of fresh blood was centrifuged and frozen for 1, 2, 3 and 4 years respectively. Then DNA was extracted and the integrity and purity of the extracted DNA were analyzed. Results Compared with fresh whole blood albuginea, the yield of frozen albuginea samples decreased significantly(P < 0.01). Compared with the alveolar membrane samples frozen for 2 years, the yield of alveolar membrane samples frozen for 1 year decreased significantly(P < 0.05). There was no significant difference in the yield of alveolar membrane samples frozen for 2, 3 and 4 years(P > 0.05). The freezing time had no significant effect on the extraction purity of samples. Freezing time had no effect on DNA integrity. Conclusion The-80 ℃ stored circumstance has not damaged the purification as well as the integrity of blood DNA except caused decreasing of its yield.
Objective To investigate the effect of blood retention time on the quality of DNA extraction of blood. Methods The supernatant of fresh blood was centrifuged and frozen for 1, 2, 3 and 4 years respectively. Then DNA was extracted and the integrity and purity of the extracted DNA were analyzed. Results Compared with fresh whole blood albuginea, the yield of frozen albuginea samples decreased significantly(P < 0.01). Compared with the alveolar membrane samples frozen for 2 years, the yield of alveolar membrane samples frozen for 1 year decreased significantly(P < 0.05). There was no significant difference in the yield of alveolar membrane samples frozen for 2, 3 and 4 years(P > 0.05). The freezing time had no significant effect on the extraction purity of samples. Freezing time had no effect on DNA integrity. Conclusion The-80 ℃ stored circumstance has not damaged the purification as well as the integrity of blood DNA except caused decreasing of its yield.
引文
[1]解晨婧.临床血液生化检验标本分析过程中影响检验结果准确性的因素探讨[J].基层医学论坛,2016,20(23):3251-3252.
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[7]吴燕琴.全血标本保存时间和温度对血常规检测结果的影响分析[J].中外医学研究,2018,16(1):62-63.
[8]陈嫣红.不同保存时间及温度对全血标本血常规检测结果的影响分析[J].中国医疗器械信息,2016,22(10):17-18.
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[11]M Shabihkhani,GM Lucey,B Wei,et al.The procurement,storage,and quality assurance of frozen blood and tissue biospecimens in pathology,biorepository,and biobank settings[J].Clin Biochem,2014,47(4-5):258-266.
[12]A Psifidi,CI Dova,G Bramis,et al.DNA Extraction Suitable for Large-Scale Whole-Genome Genotyping and Long-Term DNABanking Using Blood Samples[J].PLoS One,2015,10(1):e0115960.
[2]刘婷.血液标本存放时间对血凝中PT、APTT检测结果影响[J].世界最新医学信息文摘(连续型电子期刊),2018,18(1):137-137.
[3]刘根斌,陈端英,何孙本.评价温度和时间对血凝四项PT、FIB、APTT和TT结果的影响研究[J].现代预防医学,2016,43(5):887-890.
[4]韩海斌,周晓蓉,高书晶,等.样本保存方法及冷冻胁迫对蝗虫基因组DNA提取的影响[J].中国植保导刊,2016,37(2):18-21.
[5]肖芹,屠云洁,束婧婷,等.不同介质冷冻保存鸡血样基因组DNA提取效果比较[J].中国家禽,2018,40(9):67-69.
[6]艾卫军,彭金兰,徐霞.全血标本保存时间和温度对血常规检测结果的影响分析[J].国际检验医学杂志,2015,36(9):1293-1296.
[7]吴燕琴.全血标本保存时间和温度对血常规检测结果的影响分析[J].中外医学研究,2018,16(1):62-63.
[8]陈嫣红.不同保存时间及温度对全血标本血常规检测结果的影响分析[J].中国医疗器械信息,2016,22(10):17-18.
[9]李宁,曾光群,刘昊天,等.全血标本保存时间对血常规结果的影响[J].中国现代医生,2015,53(13):94-95,99.
[10]Bulla A,De WB,Ammerlaan W,et al.Blood DNA Yield but Not Integrity or Methylation Is Impacted After Long-Term Storage[J].Biopreservation&Biobanking,2016,14(1):29.
[11]M Shabihkhani,GM Lucey,B Wei,et al.The procurement,storage,and quality assurance of frozen blood and tissue biospecimens in pathology,biorepository,and biobank settings[J].Clin Biochem,2014,47(4-5):258-266.
[12]A Psifidi,CI Dova,G Bramis,et al.DNA Extraction Suitable for Large-Scale Whole-Genome Genotyping and Long-Term DNABanking Using Blood Samples[J].PLoS One,2015,10(1):e0115960.