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使用FTA卡存储细针穿刺样本中的乳腺癌核酸。

Use of FTA cards for the storage of breast carcinoma nucleic acid on fine-needle aspiration samples.

作者信息

Peluso Anna Lucia, Cascone Anna Maria, Lucchese Lucrezia, Cozzolino Immacolata, Ieni Antonio, Mignogna Chiara, Pepe Stefano, Zeppa Pio

机构信息

Department of Medicine and Surgery, University of Salerno, Salerno, Italy.

Department of Public Health, University of Naples "Federico II", Naples, Italy.

出版信息

Cancer Cytopathol. 2015 Oct;123(10):582-92. doi: 10.1002/cncy.21577. Epub 2015 Jun 29.

Abstract

BACKGROUND

The preservation and storage of nucleic acids is important for DNA molecular techniques. The material obtained by fine-needle aspiration (FNA) is often scanty and can not be wasted. FTA cards are filter papers that immobilize and stabilize nucleic acids and can be stored at room temperature. The current study evaluated whether nucleic acids of breast carcinoma cells, obtained by FNA in a clinical setting, may be collected, stored, and preserved on FTA cards.

METHODS

Thirty breast carcinoma, 5 non-Hodgkin lymphoma (NHL), and 5 benign reactive lymph node (RLN) cell samples obtained by FNA were stored at -80 °C and on FTA cards. DNA extraction and polymerase chain reaction were performed on cells at -80 °C and on 2 punched disks of FTA cards. Fifty nanograms of extracted DNA from both sample types were used to amplify the Janus Kinase 2 (JAK2) gene.

RESULTS

The mean value of DNA extracted from breast carcinoma cells was 28.19 ng/µL for that stored at -80 °C and 3.28 ng/µL for that stored on FTA cards. Agarose gel analysis demonstrated expected bands of DNA in 29 cases (97%) with both methods. The mean value of DNA extracted from NHL and RLN samples was 37.54 ng/µL and 4.28 ng/µL, respectively, and agarose gel analysis demonstrated bands of high molecular weight DNA in both methods. Significant differences in DNA yield were found between storage at -80 °C and FTA cards (P<.0001), but no differences were detected between 260/280 nm ratios in breast carcinoma and NHL/RLN samples.

CONCLUSIONS

FTA cards can be conveniently used for the storage of breast carcinoma cells obtained by FNA, thus providing a reliable alternative to traditional methods.

摘要

背景

核酸的保存和储存对于DNA分子技术至关重要。细针穿刺抽吸(FNA)获得的材料通常很少,不能浪费。FTA卡是一种滤纸,可固定并稳定核酸,且能在室温下储存。本研究评估了在临床环境中通过FNA获得的乳腺癌细胞核酸是否可以收集、储存并保存在FTA卡上。

方法

将通过FNA获得的30份乳腺癌、5份非霍奇金淋巴瘤(NHL)和5份良性反应性淋巴结(RLN)细胞样本分别储存在-80°C和FTA卡上。对储存在-80°C的细胞以及FTA卡的2个打孔圆盘进行DNA提取和聚合酶链反应。两种样本类型提取的50纳克DNA用于扩增Janus激酶2(JAK2)基因。

结果

储存在-80°C的乳腺癌细胞提取的DNA平均值为28.19纳克/微升,储存在FTA卡上的为3.28纳克/微升。琼脂糖凝胶分析显示,两种方法在29例(97%)中均出现了预期的DNA条带。从NHL和RLN样本中提取的DNA平均值分别为37.54纳克/微升和4.28纳克/微升,琼脂糖凝胶分析显示两种方法均出现了高分子量DNA条带。在-80°C储存和FTA卡储存之间发现DNA产量存在显著差异(P<.0001),但在乳腺癌和NHL/RLN样本的260/280纳米比值之间未检测到差异。

结论

FTA卡可方便地用于储存通过FNA获得的乳腺癌细胞,从而为传统方法提供了一种可靠的替代方法。

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