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伊拉斯姆斯医学中心组织库通过基于RNA完整性数的质量控制保证实现符合目的的冷冻组织收集。

Fit for purpose frozen tissue collections by RNA integrity number-based quality control assurance at the Erasmus MC tissue bank.

作者信息

Kap Marcel, Oomen Monique, Arshad Shazia, de Jong Bas, Riegman Peter

机构信息

Department of Pathology, Erasmus Medical Center , Rotterdam, The Netherlands .

出版信息

Biopreserv Biobank. 2014 Apr;12(2):81-90. doi: 10.1089/bio.2013.0051. Epub 2014 Feb 14.

Abstract

About 5000 frozen tissue samples are collected each year by the Erasmus Medical Center tissue bank. Two percent of these samples are randomly selected annually for RNA isolation and RNA Integrity Number (RIN) measurement. A similar quality assessment was conducted during centralization of a 20-year-old tissue collection from the cancer institute, a 15-year-old liver sample archive (-80°C), and a 13-year-old clinical pathology frozen biopsy archive (Liquid Nitrogen). Samples were divided into either high-quality (RIN ≥6.5) or low-quality overall categories, or into four "fit-for-purpose" quality groups: RIN <5: not reliable for demanding downstream analysis; 5 ≤RIN <6: suitable for RT-qPCR; 6 ≤RIN <8: suitable for gene array analysis; and RIN ≥8: suitable for all downstream techniques. In general, low RIN values were correlated with fatty, fibrous, pancreatic, or necrotic tissue. When the percentage of samples with RIN ≥6.5 is higher than 90%, the tissue bank performance is adequate. The annual 2011 quality control assessment showed that 90.3% (n=93) of all samples had acceptable RIN values; 97.4% (n=39) of the cancer institute collection had RIN values above 6.5; and 88.6% (n=123) of samples from the liver sample archive collection had RIN values higher than 6.5. As the clinical pathology biopsy collection contained only 58.8% (n=24) acceptable samples, the procurement protocols used for these samples needed immediate evaluation. When the distribution of RIN values of the different collections were compared, no significant differences were found, despite differences in average storage time and temperature. According to the principle of "fit-for-purpose" distribution, the vast majority of samples are considered good enough for most downstream techniques. In conclusion, an annual tissue bank quality control procedure provides useful information on tissue sample quality and sheds light on where and if improvements need to be made.

摘要

伊拉斯姆斯医学中心组织库每年收集约5000份冷冻组织样本。每年会从这些样本中随机抽取2%用于RNA分离和RNA完整性数值(RIN)测量。在集中整合癌症研究所20年的组织样本、一个15年的肝脏样本库(-80°C)以及一个13年的临床病理冷冻活检样本库(液氮保存)期间,进行了类似的质量评估。样本被分为高质量(RIN≥6.5)或低质量总体类别,或者分为四个“适用目的”质量组:RIN<5:对于要求较高的下游分析不可靠;5≤RIN<6:适用于逆转录定量聚合酶链反应(RT-qPCR);6≤RIN<8:适用于基因芯片分析;RIN≥8:适用于所有下游技术。一般来说,低RIN值与脂肪、纤维、胰腺或坏死组织相关。当RIN≥6.5的样本百分比高于90%时,组织库的表现是合格的。2011年的年度质量控制评估显示,所有样本中有90.3%(n=93)的RIN值可接受;癌症研究所收集的样本中有97.4%(n=39)的RIN值高于6.5;肝脏样本库收集的样本中有88.6%(n=123)的RIN值高于6.5。由于临床病理活检样本中只有58.8%(n=24)的样本可接受,因此用于这些样本的采集方案需要立即评估。当比较不同样本集的RIN值分布时,尽管平均保存时间和温度存在差异,但未发现显著差异。根据“适用目的”分布原则,绝大多数样本对于大多数下游技术来说都足够好。总之,年度组织库质量控制程序提供了有关组织样本质量的有用信息,并揭示了需要在何处以及是否需要改进。

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