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全血在室温下的保存与稳定。

The protection and stabilization of whole blood at room temperature.

作者信息

Udtha Malini, Flores Rene, Sanner Jennifer, Nomie Krystle, Backes Elizabeth, Wilbers Luke, Caldwell James

机构信息

1 Department of Nursing Systems, The University of Texas Health Science Center at Houston School of Nursing , Houston, Texas.

出版信息

Biopreserv Biobank. 2014 Oct;12(5):332-6. doi: 10.1089/bio.2014.0026.

Abstract

Preservation of biospecimens for biobanking applications traditionally involves freezing while maintaining the integrity of the product throughout multiple freeze-thaw cycles. The protection and stabilization of DNA at room temperature (RT) may eliminate the costs associated with freezer storage and reduce the maintenance costs for biobanks. However, there is a paucity of information describing the yield, purity, and integrity of DNA extracted from biospecimens stored at RT. To evaluate the yield, purity, and integrity of DNA extracted from whole blood samples stored at RT (18°C), low (-20°C), and ultra-low (-80°C) temperatures, whole blood samples from sheep and human subjects were collected, and aliquots were stored at RT (18°C), -20°C, and -80°C. Blood samples at RT were stored utilizing biostabilization technology designed to protect genomic DNA in whole blood. The quantification of the extracted DNA was determined by spectrophotometry, and the integrity was assessed following gel electrophoresis. Storage temperature did not influence the DNA yield (p=0.52); DNA yield averaged 13.6 ± 1.2 ng/μL across all storage temperatures. However, DNA yield was influenced (p=0.04) by species. The DNA yield was not influenced by a species × storage temperature interaction (p=0.84). Among the samples stored at RT, the species, type of technology utilized, and the interaction did not influence (p>0.13) DNA yield for both DNAgard and DNAstable. The 260/280 ratio was influenced by a species × storage temperature interaction (p=0.01). Generally, the 260/280 ratios were higher (p<0.05) for human samples stored at low and ultra-low temperatures compared to sheep samples stored at similar temperatures. Ambient temperature-based technologies offer an alternative to low temperature biospecimen preservation for blood that can be utilized by biobanks to reduce freezer storage costs while maintaining the quality of the biospecimen.

摘要

传统上,用于生物样本库的生物样本保存方法是冷冻,同时在多个冻融循环中保持样本的完整性。在室温(RT)下对DNA进行保护和稳定处理,可能会消除与冷冻储存相关的成本,并降低生物样本库的维护成本。然而,关于从室温保存的生物样本中提取的DNA的产量、纯度和完整性的信息却很少。为了评估从室温(18°C)、低温(-20°C)和超低温(-80°C)保存的全血样本中提取的DNA的产量、纯度和完整性,收集了绵羊和人类受试者的全血样本,并将等分样本分别储存在室温(18°C)、-20°C和-80°C下。室温下的血液样本采用旨在保护全血中基因组DNA的生物稳定技术进行储存。通过分光光度法测定提取的DNA的定量,并在凝胶电泳后评估其完整性。储存温度对DNA产量没有影响(p=0.52);所有储存温度下的DNA产量平均为13.6±1.2 ng/μL。然而,DNA产量受物种影响(p=0.04)。DNA产量不受物种×储存温度相互作用的影响(p=0.84)。在室温下储存的样本中,对于DNAgard和DNAstable,物种、所使用的技术类型以及相互作用均不影响(p>0.13)DNA产量。260/280比值受物种×储存温度相互作用的影响(p=0.01)。一般来说,与在相似温度下储存的绵羊样本相比,低温和超低温下储存的人类样本的260/280比值更高(p<0.05)。基于环境温度的技术为血液生物样本的低温保存提供了一种替代方法,生物样本库可以利用这种方法降低冷冻储存成本,同时保持生物样本的质量。

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