基于滚环扩增合成的多价适配体快速检测浆膜腔积液中肿瘤细胞的实验研究

许洁如; 陈嘉玲; 万涛; 邓红丽; 李岱容

doi:10.16016/j.1000-5404.202008006

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基于滚环扩增合成的多价适配体快速检测浆膜腔积液中肿瘤细胞的实验研究

基础医学 | 更新时间：2026-01-09

- 基于滚环扩增合成的多价适配体快速检测浆膜腔积液中肿瘤细胞的实验研究
- Rapid detection of tumor cells in serous effusion by multivalent aptamers based on rolling circle amplification
- 第三军医大学学报 2020年43卷第23期页码：2311-2319
- 作者机构：
  
  400016 重庆，重庆医科大学附属第一医院呼吸与危重症医学科
- 作者简介：
  
  [李岱容，E-mail: lidairong@hospital.cqmu.edu.cn LI Dairong, E-mail:lidairong@hospital.cqmu.edu.cn
- 基金信息：
  
  重庆市技术创新与应用发展专项面上项目(CSTC2020jscx-msxmX0063);重庆市研究生科研创新项目(CYS19187)
- DOI：10.16016/j.1000-5404.202008006
  中图分类号： R394-33;R446.9;R730.43
- 收稿：2020-08-01，
  
  修回：2020-09-23，
  
  纸质出版：2020-12-15
- 稿件说明：
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许洁如, 陈嘉玲, 万涛, 等. 基于滚环扩增合成的多价适配体快速检测浆膜腔积液中肿瘤细胞的实验研究[J]. 第三军医大学学报, 2020,43(23):2311-2319.

Jieru XU, Jialing CHEN, Tao WAN, et al. Rapid detection of tumor cells in serous effusion by multivalent aptamers based on rolling circle amplification[J]. Journal of Third Military Medical University, 2020, 43(23): 2311-2319.
许洁如, 陈嘉玲, 万涛, 等. 基于滚环扩增合成的多价适配体快速检测浆膜腔积液中肿瘤细胞的实验研究[J]. 第三军医大学学报, 2020,43(23):2311-2319. DOI： 10.16016/j.1000-5404.202008006.

Jieru XU, Jialing CHEN, Tao WAN, et al. Rapid detection of tumor cells in serous effusion by multivalent aptamers based on rolling circle amplification[J]. Journal of Third Military Medical University, 2020, 43(23): 2311-2319. DOI： 10.16016/j.1000-5404.202008006.

摘要

目的

基于多价适配体构建一种简单快速的肿瘤细胞的检测方法。

方法

通过滚环扩增反应形成多价适配体的长链支架(含重复的Poly T)，与靶向核仁素的AS1411适配体荧光信号链(带有Ploy A)杂交后，构建多价适配体识别探针Multi-AS1411-LS，并对其进行表征。用流式细胞和共聚焦显微镜分析Multi-AS1411-LS探针对肿瘤细胞识别的可行性，并进一步对识别条件进行了优化，最后对临床浆膜腔积液标本中的肿瘤细胞进行检测，评价其特异性和敏感性。

结果

构建的多价适配体识别探针在室温下5 min即可识别肿瘤细胞，比单价的AS1411适配体识别肿瘤细胞的荧光强度高100倍左右，且最低检测限为3个细胞。该多价适配体识别探针在临床标本肿瘤细胞检测中，能很好地区分良恶性细胞，敏感度为87.80%，特异性为93.10%。

结论

基于滚环扩增合成的多价适配体能快速检测浆膜腔积液中肿瘤细胞，操作方法简单，有较好的特异性和敏感度。

Abstract

Objective

To establish a simple and rapid method for detecting tumor cells based on multivalent aptamers.

Methods

Rolling circle amplification was used to form a long single stranded DNA scaffold with repeat sequences of poly T

then the obtained product was hybridized with the fluorescence signal chain of aptamer AS1411 (with poly A bases) which was nucleolin targeting. Then

a Multi-AS1411-LS recognition probe was constructed and characterized. The feasibility of probe Multi-AS1411-LS for cell line identification was analyzed by flow cytometry and confocal microscopy

and the recognition conditions were further optimized. Finally

the malignant tumor cells in clinically collected serous effusions were detected by our constructed probe Multi-AS1411-LS

and the specificity and sensitivity of our probe were evaluated.

Results

Our constructed probe Multi-AS1411-LS could recognize tumor cells in 5 min at room temperature

with the mean fluorescence intensity of cells higher by about 100 times than that of monovalent formats

and the minimum detection limit of 3 cells in 500 μL samples. The probe distinguished benign and malignant cells in 70 clinical specimens

with a sensitivity of 87.80% and a specificity of 93.10%.

Conclusion

This detection method is rapid and simple to operate

and has good specificity and sensitivity in the detection of malignant tumor cells in clinical practice.

关键词

Keywords

references

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