Suppr超能文献

从福尔马林固定石蜡包埋组织中分离RNA的石蜡包埋RNA指标(PERM)

The paraffin-embedded RNA metric (PERM) for RNA isolated from formalin-fixed, paraffin-embedded tissue.

作者信息

Chung Joon-Yong, Cho Hanbyoul, Hewitt Stephen M

机构信息

Experimental Pathology Laboratory, Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD.

Department of Obstetrics and Gynecology, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea.

出版信息

Biotechniques. 2016 May 1;60(5):239-44. doi: 10.2144/000114415. eCollection 2016.

Abstract

RNA isolated from formalin-fixed, paraffin-embedded (FFPE) tissue is commonly evaluated in both investigative and diagnostic pathology. However, the quality of the data is directly impacted by RNA quality. The RNA integrity number (RIN), an algorithm based on a combination of electrophoretic features, is widely applied to RNA isolated from paraffin-embedded tissue, but it is a poor indicator of the quality of that RNA. Here we describe the novel paraffin-embedded RNA metric (PERM) for quantifying the quality of RNA from FFPE tissue. The PERM is based on a formula that approximates a weighted area-under-the-curve analysis of an electropherogram of the extracted RNA. Using biochemically degraded RNAs prepared from experimentally fixed mouse kidney specimens, we demonstrate that PERM values correlate with mRNA transcript measurements determined using the QuantiGene system. Furthermore, PERM values correlate with real-time PCR data. Our results demonstrate that the PERM can be used to qualify RNA for different end-point studies and may be a valuable tool for molecular studies using RNA extracted from FFPE tissue.

摘要

从福尔马林固定、石蜡包埋(FFPE)组织中分离出的RNA常用于研究性病理学和诊断病理学。然而,数据质量直接受到RNA质量的影响。RNA完整性数(RIN)是一种基于电泳特征组合的算法,广泛应用于从石蜡包埋组织中分离出的RNA,但它并不能很好地指示该RNA的质量。在此,我们描述了一种用于量化FFPE组织RNA质量的新型石蜡包埋RNA指标(PERM)。PERM基于一个公式,该公式近似于对提取RNA的电泳图进行加权曲线下面积分析。使用从实验固定的小鼠肾脏标本制备的生化降解RNA,我们证明PERM值与使用QuantiGene系统测定的mRNA转录本测量值相关。此外,PERM值与实时PCR数据相关。我们的结果表明,PERM可用于鉴定不同终点研究的RNA,并且可能是使用从FFPE组织中提取的RNA进行分子研究的有价值工具。

相似文献

1
The paraffin-embedded RNA metric (PERM) for RNA isolated from formalin-fixed, paraffin-embedded tissue.
Biotechniques. 2016 May 1;60(5):239-44. doi: 10.2144/000114415. eCollection 2016.
3
Evaluation of the quality of RNA extracted from archival FFPE glioblastoma and epilepsy surgical samples for gene expression assays.
J Clin Pathol. 2018 Aug;71(8):695-701. doi: 10.1136/jclinpath-2017-204969. Epub 2018 Feb 20.
5
RNA and microRNA Stability in PAXgene-Fixed Paraffin-Embedded Tissue Blocks After Seven Years' Storage.
Am J Clin Pathol. 2018 Apr 25;149(6):536-547. doi: 10.1093/ajcp/aqy026.
6
Preparation of archival formalin-fixed paraffin-embedded mouse liver samples for use with the Agilent gene expression microarray platform.
J Pharmacol Toxicol Methods. 2013 Sep-Oct;68(2):260-268. doi: 10.1016/j.vascn.2013.02.008. Epub 2013 Mar 1.
7
Transcriptomic profiling of nuclei from paraformaldehyde-fixed and formalin-fixed paraffin-embedded brain tissues.
Anal Chim Acta. 2023 Nov 15;1281:341861. doi: 10.1016/j.aca.2023.341861. Epub 2023 Oct 12.
8
Detection of alpha human papillomaviruses in archival formalin-fixed, paraffin-embedded (FFPE) tissue specimens.
J Clin Virol. 2016 Mar;76 Suppl 1:S88-S97. doi: 10.1016/j.jcv.2015.10.007. Epub 2015 Oct 22.
9
Factors in tissue handling and processing that impact RNA obtained from formalin-fixed, paraffin-embedded tissue.
J Histochem Cytochem. 2008 Nov;56(11):1033-42. doi: 10.1369/jhc.2008.951863. Epub 2008 Aug 18.
10
Does vacuum centrifugal concentration reduce yield or quality of nucleic acids extracted from FFPE biospecimens?
Anal Biochem. 2019 Feb 1;566:16-19. doi: 10.1016/j.ab.2018.10.020. Epub 2018 Oct 19.

引用本文的文献

1
The Application of Guanidinium to Improve Biomolecule Quality in Fixed, Paraffin-embedded Tissue.
J Histochem Cytochem. 2023 Feb;71(2):87-101. doi: 10.1369/00221554231159451. Epub 2023 Mar 4.
2
A Novel HPLC-Based Method to Investigate on RNA after Fixation.
Int J Mol Sci. 2020 Oct 13;21(20):7540. doi: 10.3390/ijms21207540.
3
DV200 Index for Assessing RNA Integrity in Next-Generation Sequencing.
Biomed Res Int. 2020 Feb 25;2020:9349132. doi: 10.1155/2020/9349132. eCollection 2020.
4
6
Extraction-free whole transcriptome gene expression analysis of FFPE sections and histology-directed subareas of tissue.
PLoS One. 2019 Feb 22;14(2):e0212031. doi: 10.1371/journal.pone.0212031. eCollection 2019.
8
Understanding preanalytical variables and their effects on clinical biomarkers of oncology and immunotherapy.
Semin Cancer Biol. 2018 Oct;52(Pt 2):26-38. doi: 10.1016/j.semcancer.2017.12.008. Epub 2017 Dec 16.
9
Histomorphological and Molecular Assessments of the Fixation Times Comparing Formalin and Ethanol-Based Fixatives.
J Histochem Cytochem. 2018 Feb;66(2):121-135. doi: 10.1369/0022155417741467. Epub 2017 Nov 10.

本文引用的文献

1
A Buffered Alcohol-Based Fixative for Histomorphologic and Molecular Applications.
J Histochem Cytochem. 2016 Jul;64(7):425-40. doi: 10.1369/0022155416649579. Epub 2016 May 24.
2
Ultraviolet C radiation influences the robustness of RNA integrity measurement.
Electrophoresis. 2015 Sep;36(17):2072-81. doi: 10.1002/elps.201500082. Epub 2015 Jul 14.
4
An optimized RNA extraction method from archival formalin-fixed paraffin-embedded tissue.
Methods Mol Biol. 2010;611:19-27. doi: 10.1007/978-1-60327-345-9_2.
6
Factors in tissue handling and processing that impact RNA obtained from formalin-fixed, paraffin-embedded tissue.
J Histochem Cytochem. 2008 Nov;56(11):1033-42. doi: 10.1369/jhc.2008.951863. Epub 2008 Aug 18.
7
Evaluation of the branched-chain DNA assay for measurement of RNA in formalin-fixed tissues.
J Mol Diagn. 2008 Mar;10(2):169-76. doi: 10.2353/jmoldx.2008.070127. Epub 2008 Feb 14.
8
Quantification of mRNA using real-time RT-PCR.
Nat Protoc. 2006;1(3):1559-82. doi: 10.1038/nprot.2006.236.
9
RNA expression analysis of formalin-fixed paraffin-embedded tumors.
Lab Invest. 2007 Apr;87(4):383-91. doi: 10.1038/labinvest.3700529. Epub 2007 Feb 12.
10
Optimization of recovery of RNA from formalin-fixed, paraffin-embedded tissue.
Diagn Mol Pathol. 2006 Dec;15(4):229-36. doi: 10.1097/01.pdm.0000213468.91139.2d.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验