The Effect of Blood Clots on the Quality of RNA Extracted from PAXgene Blood RNA Tubes.

IF 1.6 4区生物学

Biopreservation and Biobanking Pub Date : 2024-04-01 Epub Date: 2023-08-03 DOI:10.1089/bio.2023.0001

Rong Tang, Ling Zhu, Ping Zhu, Ru Yin, Chunxia Zheng

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Abstract

Background: PAXgene^® Blood RNA tubes are routinely used in clinical research and molecular biology applications to preserve the stability of RNA in whole blood. However, in practice, blood clots are occasionally observed after blood collection and are often ignored. Currently, there are few studies on whether blood clots affect the quality of RNA extracted from these tubes. Materials and Methods: Fifteen pairs of non-clot and clot PAXgene Blood RNA tube samples (n = 30) were collected to form two matched groups from 15 patients. According to the maximum diameter (d) of the blood clot observed visually at the time of sample reception, the clot groups were divided into a small-clot group (0 cm < d < 0.5 cm) and a large-clot group (d ≥ 0.5 cm). RNA was extracted by the PAXgene Blood RNA Kit. To analyze the quality of RNA, its yield and purity were assessed by spectrophotometry, and integrity was measured by microfluidic electrophoresis. An A_260/280 ratio between 1.8 and 2.2 indicated purified RNA, and RNA integrity number (RIN) values ≥7.0 were considered to represent qualified integrity. Results: The median yields of RNA from the non-clot and clot groups were 3.84 (2.80-6.38) μg and 4.87 (2.77-8.30) μg, respectively. The median A_260/280 ratios were 2.08 (2.06-2.09) and 2.09 (2.07-2.11), whereas the median A_260/230 ratios were 1.77 (1.31-1.91) and 1.67 (1.21-1.94) in the two groups. In addition, the median RINs were 8.20 (8.00-8.40) and 7.20 (6.60-7.70), respectively. There were no significant differences in RNA yields, A_260/280, or A_260/230 between the two groups. However, the RIN value of the clot group was significantly lower compared with the non-clot group (p < 0.05), with RIN ≥7.0 found in all non-clot samples and 60% of clot samples (p < 0.05). Furthermore, in the clot groups, the small-clot samples had higher RIN values than large-clot samples (8.25 [7.75-8.75] vs. 6.90 [6.60-7.30], p < 0.001). Conclusions: The formation of large blood clots in PAXgene Blood RNA tubes will reduce the integrity of extracted RNA.

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血块对从 PAXgene 血液 RNA 管中提取的 RNA 质量的影响。

背景：PAXgene® 血液 RNA 管通常用于临床研究和分子生物学应用，以保持全血中 RNA 的稳定性。然而，在实践中，采血后偶尔会观察到血凝块，但往往被忽视。目前，有关血凝块是否会影响从这些试管中提取的 RNA 质量的研究很少。材料与方法：从 15 名患者中收集 15 对无血块和有血块的 PAXgene 血液 RNA 管样本（n = 30），形成两个匹配组。根据接收样本时肉眼观察到的血凝块最大直径（d），将血凝块组分为小血凝块组（0 cm < d d ≥ 0.5 cm）。用 PAXgene 血液 RNA 试剂盒提取 RNA。为了分析 RNA 的质量，用分光光度法评估其产量和纯度，并用微流控电泳法测定其完整性。A260/280 比率在 1.8 和 2.2 之间表示 RNA 已纯化，RNA 完整性编号（RIN）值≥7.0 则表示 RNA 完整性合格。结果非血块组和血块组的 RNA 产量中位数分别为 3.84 (2.80-6.38) μg 和 4.87 (2.77-8.30) μg。两组的 A260/280 比率中位数分别为 2.08（2.06-2.09）和 2.09（2.07-2.11），而 A260/230 比率中位数分别为 1.77（1.31-1.91）和 1.67（1.21-1.94）。此外，RIN 中位数分别为 8.20（8.00-8.40）和 7.20（6.60-7.70）。两组的 RNA 产量、A260/280 或 A260/230 均无明显差异。然而，与无血块组相比，血块组的 RIN 值明显较低（P P P 结论）：PAXgene 血液 RNA 管中大块血凝块的形成会降低提取 RNA 的完整性。

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来源期刊

Biopreservation and Biobanking Biochemistry, Genetics and Molecular Biology-General Biochemistry,Genetics and Molecular Biology

自引率

12.50%

发文量

114

期刊介绍： Biopreservation and Biobanking is the first journal to provide a unifying forum for the peer-reviewed communication of recent advances in the emerging and evolving field of biospecimen procurement, processing, preservation and banking, distribution, and use. The Journal publishes a range of original articles focusing on current challenges and problems in biopreservation, and advances in methods to address these issues related to the processing of macromolecules, cells, and tissues for research. In a new section dedicated to Emerging Markets and Technologies, the Journal highlights the emergence of new markets and technologies that are either adopting or disrupting the biobank framework as they imprint on society. The solutions presented here are anticipated to help drive innovation within the biobank community. Biopreservation and Biobanking also explores the ethical, legal, and societal considerations surrounding biobanking and biorepository operation. Ideas and practical solutions relevant to improved quality, efficiency, and sustainability of repositories, and relating to their management, operation and oversight are discussed as well.