Rong Tang, Ling Zhu, Ping Zhu, Ru Yin, Chunxia Zheng
{"title":"血块对从 PAXgene 血液 RNA 管中提取的 RNA 质量的影响。","authors":"Rong Tang, Ling Zhu, Ping Zhu, Ru Yin, Chunxia Zheng","doi":"10.1089/bio.2023.0001","DOIUrl":null,"url":null,"abstract":"<p><p><b><i>Background:</i></b> PAXgene<sup>®</sup> 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. <b><i>Materials and Methods:</i></b> Fifteen pairs of non-clot and clot PAXgene Blood RNA tube samples (<i>n</i> = 30) were collected to form two matched groups from 15 patients. According to the maximum diameter (<i>d</i>) of the blood clot observed visually at the time of sample reception, the clot groups were divided into a small-clot group (0 cm < <i>d</i> < 0.5 cm) and a large-clot group (<i>d</i> ≥ 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<sub>260/280</sub> ratio between 1.8 and 2.2 indicated purified RNA, and RNA integrity number (RIN) values ≥7.0 were considered to represent qualified integrity. <b><i>Results:</i></b> 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<sub>260/280</sub> ratios were 2.08 (2.06-2.09) and 2.09 (2.07-2.11), whereas the median A<sub>260/230</sub> 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<sub>260/280</sub>, or A<sub>260/230</sub> between the two groups. However, the RIN value of the clot group was significantly lower compared with the non-clot group (<i>p</i> < 0.05), with RIN ≥7.0 found in all non-clot samples and 60% of clot samples (<i>p</i> < 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], <i>p</i> < 0.001). <b><i>Conclusions:</i></b> The formation of large blood clots in PAXgene Blood RNA tubes will reduce the integrity of extracted RNA.</p>","PeriodicalId":55358,"journal":{"name":"Biopreservation and Biobanking","volume":" ","pages":"174-178"},"PeriodicalIF":1.6000,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"The Effect of Blood Clots on the Quality of RNA Extracted from PAXgene Blood RNA Tubes.\",\"authors\":\"Rong Tang, Ling Zhu, Ping Zhu, Ru Yin, Chunxia Zheng\",\"doi\":\"10.1089/bio.2023.0001\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b><i>Background:</i></b> PAXgene<sup>®</sup> 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. <b><i>Materials and Methods:</i></b> Fifteen pairs of non-clot and clot PAXgene Blood RNA tube samples (<i>n</i> = 30) were collected to form two matched groups from 15 patients. According to the maximum diameter (<i>d</i>) of the blood clot observed visually at the time of sample reception, the clot groups were divided into a small-clot group (0 cm < <i>d</i> < 0.5 cm) and a large-clot group (<i>d</i> ≥ 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<sub>260/280</sub> ratio between 1.8 and 2.2 indicated purified RNA, and RNA integrity number (RIN) values ≥7.0 were considered to represent qualified integrity. <b><i>Results:</i></b> 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<sub>260/280</sub> ratios were 2.08 (2.06-2.09) and 2.09 (2.07-2.11), whereas the median A<sub>260/230</sub> 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<sub>260/280</sub>, or A<sub>260/230</sub> between the two groups. However, the RIN value of the clot group was significantly lower compared with the non-clot group (<i>p</i> < 0.05), with RIN ≥7.0 found in all non-clot samples and 60% of clot samples (<i>p</i> < 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], <i>p</i> < 0.001). <b><i>Conclusions:</i></b> The formation of large blood clots in PAXgene Blood RNA tubes will reduce the integrity of extracted RNA.</p>\",\"PeriodicalId\":55358,\"journal\":{\"name\":\"Biopreservation and Biobanking\",\"volume\":\" \",\"pages\":\"174-178\"},\"PeriodicalIF\":1.6000,\"publicationDate\":\"2024-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biopreservation and Biobanking\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1089/bio.2023.0001\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2023/8/3 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biopreservation and Biobanking","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1089/bio.2023.0001","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/8/3 0:00:00","PubModel":"Epub","JCR":"","JCRName":"","Score":null,"Total":0}
The Effect of Blood Clots on the Quality of RNA Extracted from PAXgene Blood RNA Tubes.
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 A260/280 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 A260/280 ratios were 2.08 (2.06-2.09) and 2.09 (2.07-2.11), whereas the median A260/230 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, A260/280, or A260/230 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.
Biopreservation and BiobankingBiochemistry, 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.
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