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

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
    Bo Zhang, zhangbo@wh.iov.cn
  • Received Date: 14 May 2026
    Accepted Date: 17 June 2026
    Available online: 19 June 2026
  • 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.

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    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.

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    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.

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