[1]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,2020,42(23):2311-2319.
　XU Jieru,CHEN Jialing,WAN Tao,et al.Rapid detection of tumor cells in serous effusion by multivalent aptamers based on rolling circle amplification[J].J Third Mil Med Univ,2020,42(23):2311-2319.
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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(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第23期
页码:
2311-2319
栏目:
基础医学
出版日期:
2020-12-15

文章信息/Info

Title:
Rapid detection of tumor cells in serous effusion by multivalent aptamers based on rolling circle amplification
作者:
许洁如; 陈嘉玲; 万涛; 邓红丽; 李岱容
重庆医科大学附属第一医院呼吸与危重症医学科
Author(s):
 XU Jieru;  CHEN Jialing;  WAN Tao;  DENG Hongli;  LI Dairong

Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
多价适配体; 滚环扩增; AS1411; 肿瘤细胞检测
Keywords:
multivalent aptamers;  rolling circle amplification;  AS1411;  detection of tumor cells
分类号:
R394-33; R446.9; R730.43
文献标志码:
A
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:

ObjectiveTo establish a simple and rapid method for detecting tumor cells based on multivalent aptamers. MethodsRolling 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. ResultsOur 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%. ConclusionThis detection method is rapid and simple to operate, and has good specificity and sensitivity in the detection of malignant tumor cells in clinical practice.

相似文献/References:

[1]贾双荣,张可珺,陈伟,等.RCA技术检测甲型H1N1流感病毒耐药基因单碱基突变的应用研究[J].第三军医大学学报,2011,33(02):177.
　Jia Shuangrong,Zhang Kejun,Chen Wei,et al.Detection of single base mutations of influenza A H1N1 resistance gene using rolling circle amplification[J].J Third Mil Med Univ,2011,33(23):177.
[2]张江峰,张亚丽,曾照芳,等.利用滚环扩增技术快速检测结核分枝杆菌rpoB基因单碱基突变[J].第三军医大学学报,2013,35(20):2155.
　Zhang Jiangfeng,Zhang Yali,Zeng Zhaofang,et al.Detection of rpoB single base mutations in rifampin-resistant Mycobacterium tuberculosis by rolling circle amplification[J].J Third Mil Med Univ,2013,35(23):2155.

更新日期/Last Update: 2020-12-04