检验用于维持手术肺标本中高RNA质量的福尔马林固定方法。

Examining the Formalin Fixation Method for Maintaining High RNA Quality in Surgical Lung Specimens.

作者信息

Teishikata Takashi, Itoh Manabu, Okamoto Yusuke, Miyahara Naofumi, Nakashima Chiho, Takahashi Koichiro, Hiratsuka Masafumi, Kai Keita, Kamohara Keiji

机构信息

Department of Thoracic and Cardiovascular Surgery, Faculty of Medicine, Saga University, Saga, Japan.

Division of Hematology, Respiratory Medicine and Oncology, Department of Internal Medicine, Faculty of Medicine, Saga University, Saga, Japan.

出版信息

Thorac Cancer. 2025 Jan;16(2):e70005. doi: 10.1111/1759-7714.70005.

DOI:10.1111/1759-7714.70005

PMID:39853884

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11759292/

Abstract

BACKGROUND

Multiplex genetic testing is recommended when treating nonsmall cell lung cancer. A certain percentage of test failures in RNA assays owing to poor surgical specimen quality have been documented, and fixation failure is a possible cause. At our institution, sheet-like fixation is performed to reduce fixation time. This study aimed to compare the quality of RNA from resected lung cancer specimens following different fixation methods.

METHODS

Sheet-like fixation specimens (n = 15), conventional fixation specimens of the same resected lungs (n = 15), and other lung cancer specimens collected for conventional fixation and subjected to multiplex gene-panel testing (n = 22) were retrospectively examined. RNA was extracted from each specimen. RNA quality and quantity were compared, and the success rate of multiplex gene-panel testing was determined.

RESULTS

The DV value was significantly higher in RNA extracted from sheet-like fixation samples (median 47.5%, interquartile range [IQR]:40.3-51.5) compared with RNA extracted from conventionally fixed samples or conventionally fixed samples of other patient specimens (median 21%, IQR:5.3-29.8 and median 16.3%, IQR:9.5-27.1, respectively). No significant difference was observed in nucleic acid concentration. The multiplex genetic analysis success rate was 95% with conventional methods (one failure); however, it was 100% with the sheet-like fixation method.

CONCLUSION

Sheet-like fixation preserved RNA extracted from lung cancer specimens, resulting in lesser degradation than with conventional fixation.

摘要

背景

在治疗非小细胞肺癌时，建议进行多重基因检测。已有文献记载，由于手术标本质量差，RNA检测存在一定比例的失败情况，固定失败可能是原因之一。在我们机构，采用片状固定以缩短固定时间。本研究旨在比较不同固定方法后切除的肺癌标本中RNA的质量。

方法

回顾性检查片状固定标本（n = 15）、同一切除肺的传统固定标本（n = 15）以及为传统固定而收集并进行多重基因检测的其他肺癌标本（n = 22）。从每个标本中提取RNA。比较RNA的质量和数量，并确定多重基因检测的成功率。

结果

与从传统固定样本或其他患者标本的传统固定样本中提取的RNA相比，从片状固定样本中提取的RNA的DV值显著更高（中位数47.5%，四分位间距[IQR]：40.3 - 51.5）（分别为中位数21%，IQR：5.3 - 29.8和中位数16.3%，IQR：9.5 - 27.1）。核酸浓度未观察到显著差异。传统方法的多重基因分析成功率为95%（一例失败）；然而，片状固定方法的成功率为100%。