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一种血液游离核酸稳定剂的保存效果验证
引用本文:李晔,章宇宁,邢丽楠,李栋,赵小曼,刘青山,闫磊. 一种血液游离核酸稳定剂的保存效果验证[J]. 中国酿造, 2022, 41(5): 164-168. DOI: 10.11882/j.issn.0254-5071.2022.05.029
作者姓名:李晔  章宇宁  邢丽楠  李栋  赵小曼  刘青山  闫磊
作者单位:(1.北京电子科技职业学院 生物工程学院,北京 100176;2.无锡百泰克生物技术有限公司,江苏 无锡 271000)
基金项目:国家自然基金青年项目(51708005);
摘    要:该研究旨在研究一种新型稳定复合剂的配方,其目标是在全血样本的存储和运输过程中能防止来自细胞破裂释放的基因组脱氧核糖核酸(DNA)污染。使用抗凝剂、醛类、还原剂和猝灭剂为主要保护成分的稳定剂,分别用Qubit定量和Human β-actin实时荧光定量聚合酶链式反应(RT-FQPCR)检测不同保存温度和模拟运输条件下血细胞基因组DNA释放情况。结果表明,新型的复合稳定剂配方在4~37 ℃保存温度条件下,至少可以稳定保存14 d,在25 ℃、220 r/min模拟运输条件下,可以稳定保存14 d。新型的复合保护剂能够维持血液成分的稳定,抑制细胞破裂释放基因组DNA,在偏远地区游离核酸样本采样和运输方面有潜在的应用价值。

关 键 词:游离核酸  血液保存  基因组  样本采集  

Verification of preservation effect of a blood cell-free nucleic acid stabilizer
LI Ye,ZHANG Yuning,XING Linan,LI Dong,ZHAO Xiaoman,LIU Qingshan,YAN Lei. Verification of preservation effect of a blood cell-free nucleic acid stabilizer[J]. China Brewing, 2022, 41(5): 164-168. DOI: 10.11882/j.issn.0254-5071.2022.05.029
Authors:LI Ye  ZHANG Yuning  XING Linan  LI Dong  ZHAO Xiaoman  LIU Qingshan  YAN Lei
Affiliation:(1.Department of Biotechnology, Beijing Polytechnic, Beijing 100176, China; 2.Wuxi BioTeke Co., Ltd., Wuxi 271000, China)
Abstract:The aim of this study was to investigate the formulation of a novel stabilizing compound with the objective of preventing genomic deoxyribonucleic acid (DNA) contamination from cell rupture during storage and transport of whole blood samples. Using anticoagulants, aldehydes, reductants and quenchers as the main protective components of the stabilizers, and the release of genomic DNA of hemocytes under different storage temperatures and simulated transport conditions were detected by Qubit quantification and Human β-actin real-time fluorescence quantitative polymerase chain reaction (RT-FQPCR). The results showed that the new compound stabilizer could be stably stored at 4-37 ℃ for at least 14 d, and at 25 ℃, 220 r/min for simulated transportation for 14 d. The novel compound protective agent could maintain the stability of blood components and inhibit the release of genomic DNA by cell rupture, which had potential application value in the sampling and transportation of cell-free nucleic acid samples in remote areas.
Keywords:cell-free nucleic acid  blood storage  genome  specimen collection  
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