甲状腺生物样本在 -80°C 长期储存对 RNA 质量的影响及确保其适用性。
The effect of long-term -80°C storage of thyroid biospecimens on RNA quality and ensuring fitness for purpose.
作者信息
Mathieson William, Betsou Fay, Myshunina Tamara, Pushkarev Viktor, Pushkarev Vladimir, Shinkarkina Anna, Voskoboynyk Laryssa, Thomas Gerry A
机构信息
Department of Surgery and Cancer, Imperial College London, London, UK.
Integrated Biobank of Luxembourg, Luxembourg, Luxembourg.
出版信息
J Clin Pathol. 2016 Dec;69(12):1105-1108. doi: 10.1136/jclinpath-2016-203697. Epub 2016 May 27.
AIMS
To establish whether RNA degrades in long-term storage at -80°C and whether RNA integrity numbers (RINs) determine 'fitness for purpose' in severely degraded RNA.
METHODS
RNA was extracted from 549 thyroid biospecimens stored at -80°C for 0.1-10.9 years then their RINs correlated with storage time. RT-PCR for 65, 265, 534 and 942 base pair amplicons of hydroxymethylbilane synthase was used to measure amplicon length in RNA from cryopreserved and FFPE biospecimens that were equally degraded according to RIN.
RESULTS
Storage time did not correlate with RIN. Longer amplicons were obtained from cryopreserved samples than FFPE samples with equal RINs.
CONCLUSIONS
RNA does not degrade in thyroid biospecimens stored for long periods of time at -80°C. Although RINs are known to predict amenability to analytical platforms in good quality samples, this prediction is unreliable in severely degraded samples.
目的
确定RNA在-80°C长期储存时是否会降解，以及RNA完整性数值（RIN）是否能决定严重降解的RNA的“适用性”。
方法
从549份储存在-80°C达0.1 - 10.9年的甲状腺生物样本中提取RNA，然后将其RIN与储存时间进行关联分析。使用针对羟甲基胆色素原合酶65、265、534和942碱基对扩增子的逆转录聚合酶链反应（RT-PCR）来测量来自冷冻保存和福尔马林固定石蜡包埋（FFPE）生物样本中RNA的扩增子长度，这些样本根据RIN值被判定为同等程度降解。
结果
储存时间与RIN不相关。在具有相同RIN值的情况下，从冷冻保存样本中获得的扩增子比FFPE样本更长。
结论
储存在-80°C的甲状腺生物样本中的RNA不会降解。虽然已知RIN可预测高质量样本对分析平台的适用性，但在严重降解的样本中这种预测并不可靠。
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