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揭开福尔马林固定石蜡包埋存档组织块的奥秘:基因组DNA提取的宝贵资源。

Unveiling the arcanum of formalin-fixed paraffin-embedded archival tissue blocks: A valuable resource for genomic DNA extraction.

作者信息

Begum Sameera, Prabhu Vishnudas, Mohanty Varshasnata, Upadhyaya K Krishnaraj, Abdulla Riaz

机构信息

Department of Oral Pathology, Yenepoya Dental College, Yenepoya (Deemed to be University), Mangaluru, Karnataka, India.

Center for Systems Biology and Molecular Medicine, Yenepoya Research Centre, Yenepoya (Deemed to be University), Mangaluru, Karnataka, India.

出版信息

J Oral Maxillofac Pathol. 2022 Apr-Jun;26(2):289. doi: 10.4103/jomfp.jomfp_424_20. Epub 2022 Jun 28.

Abstract

BACKGROUND

Formalin-fixed paraffin-embedded (FFPE) tissue blocks are routinely preserved after pathological diagnosis and possess tremendous potential for biomarker discovery. These archival samples are prone to degradation on prolonged storage due to the formalin cross-linking.

AIMS

This study aimed to evaluate whether the storage period of the formalin-fixed paraffin-embedded tumor blocks had a significant impact on the yield and purity of the isolated DNA archived for 11 years.

SETTINGS AND DESIGN

A retrospective study was carried out in the Department of Oral Pathology and Microbiology in accordance with the Institutional Ethics Committee.

MATERIALS AND METHODS

Genomic DNA extraction was performed using TaKaRa DEXPAT Easy DNA kit from 40 FFPE tissue blocks of oral squamous cell carcinoma archived for 11 years (2006-2017). NanoDrop spectrophotometer was used to determine the DNA yield (A260) and purity (A260/A280 ratio). The quality of DNA fragments was validated using agarose gel electrophoresis.

STATISTICAL ANALYSIS USED

Statistical analysis was obtained by SPSS 22, MS Excel and analyzed using the analysis of variance (ANOVA) test. < 0.05 was set for statistical significance.

RESULTS

There was no statistically significant difference observed both in terms of DNA yield ( = 0.996) and purity ( = 0.997) of FFPE tumor blocks archived for 11 years among the study groups.

CONCLUSIONS

It was concluded that, irrespective of years of storage of the FFPE, it is possible to extract genomic DNA and use it for molecular studies.

摘要

背景

病理诊断后,福尔马林固定石蜡包埋(FFPE)组织块通常会被保存下来,在生物标志物发现方面具有巨大潜力。由于福尔马林交联作用,这些存档样本在长期储存时容易降解。

目的

本研究旨在评估福尔马林固定石蜡包埋肿瘤组织块的储存时间对保存11年的分离DNA的产量和纯度是否有显著影响。

设置与设计

在口腔病理学和微生物学系按照机构伦理委员会的要求进行了一项回顾性研究。

材料与方法

使用TaKaRa DEXPAT Easy DNA试剂盒从40个保存11年(2006 - 2017年)的口腔鳞状细胞癌FFPE组织块中提取基因组DNA。使用NanoDrop分光光度计测定DNA产量(A260)和纯度(A260/A280比值)。通过琼脂糖凝胶电泳验证DNA片段的质量。

所用统计分析方法

采用SPSS 22和MS Excel进行统计分析,并使用方差分析(ANOVA)检验。设定P < 0.05为具有统计学意义。

结果

在研究组中,保存11年的FFPE肿瘤组织块的DNA产量(P = 0.996)和纯度(P = 0.997)均未观察到统计学上的显著差异。

结论

得出的结论是,无论FFPE保存年限如何,都有可能提取基因组DNA并将其用于分子研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d55e/9364647/f095a3c76984/JOMFP-26-289a-g001.jpg

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