. doi: 10.1016/j.virs.2026.06.012
Citation: Ya-Nan Zhang, Yan-Ning Hu, Yu-Jia Lu, Xiao-Dan Li, Ana Carolina Gomes Jardim, Bo Zhang. Development of a replicon cell line-based high throughput antiviral assay for screening inhibitors of Mayaro virus .VIROLOGICA SINICA, 2026, 41(3) : 724-727.  http://dx.doi.org/10.1016/j.virs.2026.06.012

基于复制子细胞系的马亚罗病毒高通量抗病毒检测方法

  • 马亚罗病毒(MAYV)是披膜病毒科中一种再度出现的甲病毒,近年来随着美洲地区暴发相关疫情,该病毒已受到全球公共卫生机构的关注。目前,针对马亚罗病毒感染尚无特异性治疗药物及获批上市的疫苗。马亚罗病毒通常被列为生物安全3级病原,由于操作活病毒需要在高等级安全实验室内进行,极大限制了其抗病毒研究的开展。本研究构建了一种马亚罗病毒复制子,该复制子包含病毒复制酶蛋白、嘌呤霉素乙酰转移酶筛选标记,且筛选标记与海肾萤光素酶报告基因通过口蹄疫病毒2A型自剪切肽分隔。经嘌呤霉素筛选,我们获得了携带该复制子的稳定BHK-21细胞系。利用已知的甲病毒抑制剂,我们验证了该复制子细胞系用于抗病毒药物筛选的可行性。此外,通过优化细胞铺板数量、化合物浓度及孵育时间,我们建立了稳定可靠的基于复制子细胞的高通量筛选(HTS)模型,其计算所得Z′值大于0.5。该基于复制子的高通量筛选体系无需操作野生型马亚罗病毒,可为马亚罗病毒的抗病毒药物筛选提供安全的研究平台。

Development of a replicon cell line-based high throughput antiviral assay for screening inhibitors of Mayaro virus

  • Highlights
    1 Construction of a MAYV replicon expressing both PAC and Rluc genes.
    2 A stable BHK-21 cell line containing MAYV-Rep with puromycin selection was obtained.
    3 A robust replicon cell-based HTS assay based was established.

  • 加载中
    1. Amorim, R., De Meneses, M.D.F., Borges, J.C., Da Silva Pinheiro, L.C., Caldas, L.A., Cirne-Santos, C.C., De Mello, M.V.P., De Souza, A.M.T., Castro, H.C., De Palmer Paixao, I.C.N., Campos, R.M., Bergmann, I.E., Malirat, V., Bernardino, A.M.R., Rebello, M.A.,Ferreira, D.F., 2017. Thieno[2,3-b]pyridine derivatives: a new class of antiviral drugs against Mayaro virus. Arch Virol. 162, 1577-1587.

    2. Caldas, L.A., Ferreira, D.F.,Freitas, T.R.P., 2018. Prostaglandin A1 triggers Mayaro virus inhibition and heat shock protein 70 expression in an epithelial cell model. Rev Soc Bras Med Trop. 51, 584-590.

    3. Camini, F.C., Da Silva, T.F., Da Silva Caetano, C.C., Almeida, L.T., Ferraz, A.C., Alves Vitoreti, V.M., De Mello Silva, B., De Queiroz Silva, S., De Magalhaes, J.C.,De Brito Magalhaes, C.L., 2018. Antiviral activity of silymarin against Mayaro virus and protective effect in virus-induced oxidative stress. Antiviral Res. 158, 8-12.

    4. Carvalho, C.A., Sousa, I.P., Jr., Silva, J.L., Oliveira, A.C., Goncalves, R.B.,Gomes, A.M., 2014. Inhibition of Mayaro virus infection by bovine lactoferrin. Virology. 452-453, 297-302.

    5. Elshabrawy, H.A., Fan, J., Haddad, C.S., Ratia, K., Broder, C.C., Caffrey, M.,Prabhakar, B.S., 2014. Identification of a broad-spectrum antiviral small molecule against severe acute respiratory syndrome coronavirus and Ebola, Hendra, and Nipah viruses by using a novel high-throughput screening assay. J Virol. 88, 4353-4365.

    6. Li, J.Q., Deng, C.L., Gu, D., Li, X., Shi, L., He, J., Zhang, Q.Y., Zhang, B.,Ye, H.Q., 2018. Development of a replicon cell line-based high throughput antiviral assay for screening inhibitors of Zika virus. Antiviral Res. 150, 148-154.

    7. Li, Q., Maddox, C., Rasmussen, L., Hobrath, J.V.,White, L.E., 2009. Assay development and high-throughput antiviral drug screening against Bluetongue virus. Antiviral Res. 83, 267-273.

    8. Li, X., Zhang, H., Zhang, Y., Li, J., Wang, Z., Deng, C., Jardim, A.C.G., Terzian, A.C.B., Nogueira, M.L.,Zhang, B., 2019. Development of a rapid antiviral screening assay based on eGFP reporter virus of Mayaro virus. Antiviral Res. 168, 82-90.

    9. Tiong-Yip, C.L., Plant, H., Sharpe, P., Fan, J., Rich, K., Gorseth, E.,Yu, Q., 2014. Development of a high-throughput replicon assay for the identification of respiratory syncytial virus inhibitors. Antiviral Res. 101, 75-81.

    10. Zhang, J.H., Chung, T.D.,Oldenburg, K.R., 1999. A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays. J Biomol Screen. 4, 67-73.

    11. Zhang, Q.Y., Li, X.D., Liu, S.Q., Deng, C.L., Zhang, B.,Ye, H.Q., 2017. Development of a stable Japanese encephalitis virus replicon cell line for antiviral screening. Arch Virol. 162, 3417-3423.

  • 加载中

Figures(1)

Article Metrics

Article views(40) PDF downloads(1) Cited by(0)

Related
Proportional views
    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Development of a replicon cell line-based high throughput antiviral assay for screening inhibitors of Mayaro virus

      Corresponding author: Ya-Nan Zhang, ynzhang@wh.iov.cn
      Corresponding author: Bo Zhang, zhangbo@wh.iov.cn
    • a. State Key Laboratory of Virology and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China;
    • b. University of Chinese Academy of Sciences, Beijing 100049, China;
    • c. Hunan Normal University School of Medicine, Changsha 410081, China;
    • d. Laboratory of Antiviral Research, Institute of Biomedical Sciences, Federal University of Uberlândia, Uberlândia 38405-31, Brazil

    Abstract: Highlights
    1 Construction of a MAYV replicon expressing both PAC and Rluc genes.
    2 A stable BHK-21 cell line containing MAYV-Rep with puromycin selection was obtained.
    3 A robust replicon cell-based HTS assay based was established.

    Figure (1)  Reference (11) Relative (20)

    目录

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return