Impact of Cold Ischemic Time on Nucleic Acid Quality in Cytology Samples for Cancer Gene Analysis

IF 1.1 4区医学 Q4 CELL BIOLOGY

Cytopathology Pub Date : 2025-04-01 DOI:10.1111/cyt.13491

Hideyuki Abe, Akihiko Kawahara, Ryou Makino, Takato Kumagae, Jun Akiba, Hironori Kusano

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引用次数: 0

Abstract

Objective

Despite the growing attention on the use of cytology samples for genetic analysis, the impact of the time from sample collection to fixation, referred to as ‘cold’ ischemic time, has not been sufficiently studied. Therefore, we investigated the quality of nucleic acids prior to fixation in body cavity fluid samples, focusing on the benign/malignant status, differences in collection methods and tumour cell content.

Methods

We analysed 49 body cavity fluid samples collected using different methods: aspiration (n = 21), drainage (n = 5), and surgery (n = 15). The samples were collected from 26 malignant and 23 benign cases. DNA and RNA were extracted from all samples, and their yield and quality were assessed using Agilent TapeStation. Tumour cell content was calculated as the ratio of malignant to benign cells.

Results

DNA and RNA yields were significantly higher in malignant than in benign cases (p < 0.05). Regarding nucleic acid quality, there was a significant difference in RNA quality between malignant and benign cases, but no significant difference in DNA quality. Among the 26 malignant cases, there were significant differences in integrity number (RIN) and RNA percentage of nucleic acid fragments with > 200 nucleotides (DV200) between samples collected via aspiration and surgery (p < 0.05). Tumour cell content (median, 36%; range, 20%–72%) showed no correlation with nucleic acid yield or quality.

Conclusions

This study's findings suggest that DNA extracted from cytology samples is highly stable, while RNA is affected by cold ischemic time. Thus, prompt fixation after collection is necessary to maintain RNA quality.

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冷缺血时间对肿瘤基因分析细胞学样品核酸质量的影响。

目的：尽管人们越来越关注使用细胞学样本进行遗传分析，但从样本收集到固定的时间（称为“冷”缺血时间）的影响尚未得到充分研究。因此，我们对体腔液标本固定前的核酸质量进行了调查，重点关注其良/恶性状态、采集方法的差异和肿瘤细胞含量。方法：对49份体腔液标本进行分析，分别采用抽吸（n = 21）、引流（n = 5）和手术（n = 15）。恶性肿瘤26例，良性肿瘤23例。从所有样品中提取DNA和RNA，并使用Agilent tapstation评估其产量和质量。肿瘤细胞含量计算为恶性细胞与良性细胞的比值。结果：恶性肿瘤中DNA和RNA的产率明显高于良性肿瘤（p200核苷酸（DV200））。结论：本研究结果提示细胞学样本中提取的DNA具有高度稳定性，而RNA受冷缺血时间的影响。因此，收集后及时固定是保持RNA质量的必要条件。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cytopathology 生物-病理学

CiteScore

2.30

自引率

15.40%

发文量

107

审稿时长

6-12 weeks

期刊介绍： The aim of Cytopathology is to publish articles relating to those aspects of cytology which will increase our knowledge and understanding of the aetiology, diagnosis and management of human disease. It contains original articles and critical reviews on all aspects of clinical cytology in its broadest sense, including: gynaecological and non-gynaecological cytology; fine needle aspiration and screening strategy. Cytopathology welcomes papers and articles on: ultrastructural, histochemical and immunocytochemical studies of the cell; quantitative cytology and DNA hybridization as applied to cytological material.