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Volume 35 Issue 3
June 2020

    Zhiqiang Chen, Qihan Wu, Jing Chen, Xiaohua Ni and Jianfeng Dai. A DNA Aptamer Based Method for Detection of SARS-CoV-2 Nucleocapsid Protein[J]. Virologica Sinica, 2020, 35(3): 351-354. doi: 10.1007/s12250-020-00236-z
    Citation: Zhiqiang Chen, Qihan Wu, Jing Chen, Xiaohua Ni, Jianfeng Dai. A DNA Aptamer Based Method for Detection of SARS-CoV-2 Nucleocapsid Protein .VIROLOGICA SINICA, 2020, 35(3) : 351-354.  http://dx.doi.org/10.1007/s12250-020-00236-z
    shu

    一种基于DNA核酸适配体的SRAS-CoV-2核衣壳蛋白检测方法

    cstr: 32224.14.s12250-020-00236-z
    • 1.

      苏州大学生物医学研究院

    • 2.

      苏州大学附属第一医院核医学科

    • 3.

      复旦大学生殖调控卫健委重点实验室，上海计划生育科学研究所

    • 通讯作者： 倪晓华, xhni_sippr@163.com, ORCID: http://orcid.org/0000-0002-9527-6426
      ; 戴建锋, daijianfeng@suda.edu.cn, ORCID: http://orcid.org/0000-0002-1871-8390
    • 收稿日期： 2020-04-16
      录用日期： 2020-04-30
      出版日期： 2020-05-25
    • 新型冠状病毒SARS-CoV-2引起的人类疾病COVID-19正在全世界范围内大流行。感染人群的快速检测和隔离治疗，是控制疾病的关键。目前病毒的诊断主要是基于核酸和抗体的检测方法。根据2003年SARS疾病流行期间的经验，SARS-CoV的核衣壳蛋白（Nucleocapsid（N）protein）是病毒感染的一个早期诊断指标。本文基于SARS-CoV-2和SARS-CoV的N蛋白的高度同源性，利用针对SARS-CoV N蛋白的DNA单链核酸适配体，设计了一系列识别SARS-CoV-2的核酸适配体。这些核酸适配体能够特异性的识别SARS-CoV-2 N蛋白，并能检测到稀释于人血清中的重组N蛋白。N蛋白的核酸适配体还可以在蛋白印迹实验中特异性检测到SARS-CoV-2 N蛋白，与抗N蛋白抗体具有类似的灵敏度。因此，本论文中设计的DNA核酸适配体可以用于SARS-CoV-2 N蛋白的快速检测。

    A DNA Aptamer Based Method for Detection of SARS-CoV-2 Nucleocapsid Protein

    • 1.

      Institutes of Biology and Medical Sciences, Jiangsu Key Laboratory of Infection and Immunity, Soochow University, Suzhou 215123, China

    • 2.

      Department of Nuclear Medicine, The First Affiliated Hospital of Soochow University, Suzhou 215006, China

    • 3.

      NHC Key Lab of Reproduction Regulation (Shanghai Institute of Planned Parenthood Research), Fudan University, Shanghai 200035, China

    • Corresponding author: Xiaohua Ni, xhni_sippr@163.com Jianfeng Dai, daijianfeng@suda.edu.cn
    • ORCID: http://orcid.org/0000-0002-9527-6426; http://orcid.org/0000-0002-1871-8390
    • Received Date: 16 April 2020
      Accepted Date: 30 April 2020
      Published Date: 25 May 2020
    • Altogether, we have identified a novel method for the detection of SRAS-CoV-2 N protein using DNA based aptamers. Although the aptamers used in this study were designed based on an aptamer previously selected for SARS-CoV N protein, they bind to SRAS-CoV-2 N protein with a high affinity. Most importantly, SARS-CoV-2 N protein shares very low similarity (16%–38%) with N protein from other five known human coronaviruses except for SARS-CoV. Because there are no SARS-CoV cases reported since 2004, our aptamer-based method can be used as a supplementation to the current diagnosis of SARS-CoV-2.
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      4. Chen HL, Hsiao WH, Lee HC, Wu SC, Cheng JW (2015) Selection and characterization of DNA aptamers targeting all four serotypes of dengue viruses. PLoS ONE 10:e0131240
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      5. Cho SJ, Woo HM, Kim KS, Oh JW, Jeong YJ (2011) Novel system for detecting SARS coronavirus nucleocapsid protein using an ssDNA aptamer. J Biosci Bioeng 112:535–540
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      6. Jang KJ, Lee NR, Yeo WS, Jeong YJ, Kim DE (2008) Isolation of inhibitory RNA aptamers against severe acute respiratory syndrome (SARS) coronavirus NTPase/Helicase. Biochem Biophys Res Commun 366:738–744
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      8. Lee KH, Zeng H (2017) Aptamer-Based ELISA Assay for Highly Specific and Sensitive Detection of Zika NS1 Protein. Anal Chem 89:12743–12748
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      13. Wu D, Wu T, Liu Q, Yang Z (2020) The SARS-CoV-2 outbreak: what we know. Int J Infect Dis. https://doi.org/10.1016/j.ijid.2020.03.004
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    • Electronic Supplementary Material

      10.1007s12250-020-00236-z-ESM.pdf
      10.1007s12250-020-00236-z-ESM1.pdf

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      A DNA Aptamer Based Method for Detection of SARS-CoV-2 Nucleocapsid Protein

        Corresponding author: Xiaohua Ni, xhni_sippr@163.com
        Corresponding author: Jianfeng Dai, daijianfeng@suda.edu.cn
      • 1. Institutes of Biology and Medical Sciences, Jiangsu Key Laboratory of Infection and Immunity, Soochow University, Suzhou 215123, China
      • 2. Department of Nuclear Medicine, The First Affiliated Hospital of Soochow University, Suzhou 215006, China
      • 3. NHC Key Lab of Reproduction Regulation (Shanghai Institute of Planned Parenthood Research), Fudan University, Shanghai 200035, China
      • Received Date: 16 April 2020
      • Accepted Date: 30 April 2020
      • Published Date: 25 May 2020

      Abstract: Altogether, we have identified a novel method for the detection of SRAS-CoV-2 N protein using DNA based aptamers. Although the aptamers used in this study were designed based on an aptamer previously selected for SARS-CoV N protein, they bind to SRAS-CoV-2 N protein with a high affinity. Most importantly, SARS-CoV-2 N protein shares very low similarity (16%–38%) with N protein from other five known human coronaviruses except for SARS-CoV. Because there are no SARS-CoV cases reported since 2004, our aptamer-based method can be used as a supplementation to the current diagnosis of SARS-CoV-2.

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