High-Sensitive Detection of Small-Cell Lung Cancer Cells Based on Terminal Deoxynucleotidyl Transferase-Mediated Extension Polymerization Aptamer Probe

作者全名："Chen, Jialing; Xu, Jieru; Wan, Tao; Deng, Hongli; Li, Dairong"

作者地址："[Chen, Jialing; Xu, Jieru; Wan, Tao; Deng, Hongli; Li, Dairong] Chongqing Med Univ, Affiliated Hosp 1, Dept Resp & Crit Care Med, Chongqing 400016, Peoples R China"

通信作者："Li, DR (corresponding author), Chongqing Med Univ, Affiliated Hosp 1, Dept Resp & Crit Care Med, Chongqing 400016, Peoples R China."

来源：ACS BIOMATERIALS SCIENCE & ENGINEERING

ESI学科分类：MATERIALS SCIENCE

WOS号：WOS:000627588400030

JCR分区：Q2

影响因子：5.8

年份：2021

卷号：7

期号：3

开始页：1169

结束页：1180

文献类型：Article

关键词：small-cell lung cancer; aptamer; terminal deoxynucleotidyl transferase; signal amplification

摘要："Small-cell lung cancer (SCLC) is characterized by early metastasis and high invasiveness, poor prognosis, and a low five-year survival rate. Therefore, the development of the effective detection of SCLC cells and imaging methods has potential significance for the prognosis and treatment of SCLC. We designed a terminal deoxynucleotidyl transferase (TdT)-mediated extension polymerization aptamer probe (denoted as TEPAP). Aptamer HCC03 was used as an element of recognizing SCLC, and it was extended as a long poly(T) tail at the 3'-hydroxyl terminus by TdT and then hybridized with short poly(A) labeled with 6-carboxyfluorescein (FAM) to construct TEPAP for the high-sensitivity detection of SCLC. The results showed that the probe could specifically recognize NCI-H446 cells. Compared with HCC03 labeled with FAM, TEPAP has demonstrated a higher fluorescence signal in recognizing NCI-H446 cells, and the fluorescence intensity of TEPAP recognizing the target cells was 10 times higher than that of nontarget cells. Flow cytometric analysis showed that the detection limit of this method was as low as 17 NCI-H446 cells in 200 mu L of binding buffer. In the application of clinical cytology cell blocks, the sensitivity, specificity, and accuracy of TEPAP were 89.74, 94.44, and 91.23%, respectively. The high sensitivity and specificity of TEPAP in the application of clinical samples show that the proposed probe has great potential in the diagnosis of SCLC."

基金机构：Program of Chongqing Special Social Livelihood of Science and Technology Innovation [cstc2020 jscx-msxmX0063]; Graduate Scientific Research and Innovation Project of Chongqing [CYS19187]; Research Project of Innovative Talents Training Project for Primary and Middle School students of Chongqing [CY200402]

基金资助正文："This work was supported by the Program of Chongqing Special Social Livelihood of Science and Technology Innovation (NO. cstc2020 jscx-msxmX0063), the Graduate Scientific Research and Innovation Project of Chongqing (NO. CYS19187), and the Research Project of Innovative Talents Training Project for Primary and Middle School students of Chongqing (CY200402)."