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一项生物样本库计划实施后儿科急性白血病病例核酸产量的质量和数量审计

Audit of Quality and Quantity of Nucleic Acid Yield from Pediatric Acute Leukemia Cases Following a Bio-banking Initiative.

作者信息

Kumar A, Singh M, Bhatia P, Singh A

机构信息

1University of Pittsburgh School of Medicine, Pittsburgh, USA.

2Pediatric Hematology - Oncology Unit, Department of Pediatrics, APC, PGIMER, Chandigarh, India.

出版信息

Indian J Hematol Blood Transfus. 2019 Jan;35(1):77-82. doi: 10.1007/s12288-018-0975-4. Epub 2018 Jun 21.

Abstract

Information which can be harvested from a biological sample has greatly improved with advancements in diagnostic technologies. However, in developing countries, the awareness about usefulness of bio-banking concept is lacking and centres which do offer it, depend mainly on - 20 or - 80 °C for sample storage due to lack of sophisticated infrastructure like vapour phase nitrogen storage preservation. Hence in these resource constraint settings, timely audit of quality of nucleic acids extractable from samples stored is of utmost importance. In this study, we explore the effect of - 20 °C storage over nucleic acids (DNA/RNA) isolated from blood samples of 180 patients with various leukaemia's following a bio-banking initiative. We observed that the integrity and quality of both DNA and RNA were maintained in 70 and 80% samples respectively over time as reflected by their concentration measurements and inherent uniform expression of housekeeping gene GAPDH. Only 3.7% of the RNA samples and 4.2% of the DNA samples yielded very low concentrations despite minimizing processing and technical loss. In nutshell, audit of our biobank sample yield highlights that storage of blood samples at - 20 °C does not compromise the fidelity of nucleic acids for future diagnostic and research work in a resource constraint setting.

摘要

随着诊断技术的进步,从生物样本中获取的信息有了极大改善。然而,在发展中国家,人们对生物样本库概念的有用性缺乏认识,而那些提供生物样本库服务的中心,由于缺乏诸如气相氮储存保存等先进基础设施,主要依赖-20℃或-80℃进行样本储存。因此,在这些资源有限的环境中,及时对储存样本中可提取核酸的质量进行评估至关重要。在本研究中,我们通过一项生物样本库计划,探讨了-20℃储存对180例不同白血病患者血液样本中分离出的核酸(DNA/RNA)的影响。我们观察到,随着时间推移,分别有70%和80%的样本中DNA和RNA的完整性和质量得以保持,这通过它们的浓度测量以及管家基因GAPDH的固有均匀表达得以体现。尽管尽量减少了处理和技术损失,但只有3.7%的RNA样本和4.2%的DNA样本产生了极低的浓度。简而言之,我们对生物样本库样本产量的评估表明,在资源有限的环境中,将血液样本储存在-20℃不会影响核酸的保真度,以用于未来的诊断和研究工作。

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