Skip to main content

Advertisement

SpringerLink
Log in

Development of a sandwich ELISA for the detection of Chinese sacbrood virus infection

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

Chinese sacbrood disease (CSBD) is a highly pathogenic infectious disease in bees that is caused by Chinese sacbrood virus (CSBV). Although several molecular detection methods have been developed for CSBV, there are no commercially available enzyme-linked immunosorbent assay (ELISA) kits. We therefore developed a sandwich ELISA to detect CSBV antigens. To this end, monoclonal antibodies were produced using VP2 as an immunogen and subsequently characterized. Hybridomas were screened for the secretion of immunoglobulin G (IgG). Using an unlabeled monoclonal antibody (mAb) for coating and a horseradish peroxidase (HRP)-labeled mAb for detection, a CSBV sandwich ELISA method was established. This method showed specificity for CSBV and did not show cross-reactivity with other bee viruses. The detection limit of the sandwich ELISA was 3.675 × 104 copies/µL. Sixty bee larvae were tested using our sandwich ELISA method, and the presence of CSBV was verified by reverse transcription polymerase chain reaction (RT-PCR). The total coincidence rate was 90%. Thus, a sandwich ELISA method with high specificity and accuracy and a detection limit of 3.675 × 104 copies/µL has been successfully developed and can be used for the clinical detection of CSBV. This method will support rapid diagnosis, real-time monitoring, and early warning of CSBD.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Ma MX (2014) New insights of sacbrood virus. Virologica Sinica 29(6):410–413

    Article  Google Scholar 

  2. Berényi O, Bakonyi T, Derakhshifar I, Köglberger H, Nowotny N (2006) Occurrence of six honeybee viruses in diseased austrian apiaries. Appl Environ Microbiol 72(4):2414–2420

    Article  Google Scholar 

  3. Hassanyar AK, Huang S, Li Z et al (2019) Prevalence of bee viruses in Apis cerana cerana population from different locations in the Fujian Province of China. Microbiol Open 8(9):e00830

    Article  Google Scholar 

  4. Li M, Fei D, Sun L, Ma M (2019) Genetic and phylogenetic analysis of Chinese sacbrood virus isolates from Apis mellifera. Peer J 7:e8003

    Article  Google Scholar 

  5. Sun L, Li M, Fei D, Hu Y, Ma M (2017) Chinese sacbrood virus infection in Apis mellifera, Shandong, China, 2016. Virus Res 242:96–99

    Article  CAS  Google Scholar 

  6. Ma MX, Li M, Cheng J, Yang S, Wang SD, Li PF (2011) Molecular and biological characterization of Chinese sacbrood virus LN isolate. Comp Funct Genom 2011:409386

    Article  Google Scholar 

  7. Ma MX, Ma C, Cheng J, Xie Z, Li M, Fei DL (2011) Prediction and identification of structural proteins of chinese sacbrood virus LN isolate. Chem J Chin Univ 32(8):1768–1773

    CAS  Google Scholar 

  8. Fei DL, Zhang H, Diao QY, Jiang LL, Wang Q, Zhong Y, Fan ZB, Ma MX (2015) Correction: codon optimization, expression in escherichiacoli, and immunogenicity of recombinant Chinese Sacbrood virus (CSBV) structural proteins VP1, VP2, and VP3. PloS One 10:e0128486

    Article  Google Scholar 

  9. Li FB, Fei DL, Zhang H, Hu Y, Wei D, Zhang H, Yue JJ, Ma MX (2017) Optimized expression of the VP2 code of the structural protein of CSBV and its immunogenicity. Chin J Virol 02:108–114

    Google Scholar 

  10. Zhang XY, Fei DL, Sun L, Li M, Ma YY, Wang C, Huang SC, Ma MX (2019) Identification of the novel host protein interacting with the structural protein vp1 of chinese sacbrood virus by yeast two-hybrid screening. Front Microbiol 10:2192

    Article  Google Scholar 

  11. Fei DL, Wei D, Yu XL, Yue JJ, Li M, Sun L, Jiang LL, Li Y, Diao QY, Ma MX (2018) Screening of binding proteins that interact with chinese sacbrood virus vp3 capsid protein in, apis cerana, larvae cdna library by the yeast two-hybrid method. Virus Res 248:24–30

    Article  CAS  Google Scholar 

  12. Xia X, Zhou B, Wei T (2015) Complete genome of Chinese sacbrood virus from Apis cerana and analysis of the 3C-like cysteine protease. Virus Genes 50(2):277–285

    Article  CAS  Google Scholar 

  13. Zhang J, Feng J, Liang Y, Zhang Q, Lu X, Chen D, Zhou Z (2001) Three-dimensional structure of the Chinese Sacbrood bee virus. Sci China C Life Sci 44(4):443–448

    Article  CAS  Google Scholar 

  14. Zhang H (2018) Preparation of monoclonal antibody against Saccharomyces cerevisiae larvirus and establishment of agar diffusion assay. Dissertation, Jinzhou Medical University

  15. Mingxiao M, Jinhua L, Yingjin S, Li L, Yongfei L (2013) TaqMan MGB probe fluorescence real-time quantitative PCR for rapid detection of Chinese Sacbrood virus. PLoS One 8(2):e52670

    Article  Google Scholar 

  16. Ma MX, Li M, Yuan CY (2010) Development of a rt-pcr method for determination of chinese sacbrood virus. Chin J Biol 23(4):425–427

    CAS  Google Scholar 

  17. Song Z, Wang Z, Feng X, Liu J, Wei C, Cai Y, Meng Q, Zhou L (2010) A real-time TaqMan PCR assay to detect Sacbrood virus in honeybee and honeybee peoducts. Acta Entomol Sinica 53(8):920–925

    CAS  Google Scholar 

  18. Ma MX, Ma C, Li M, Wang S, Yang S, Wang S (2011) Loop-mediated isothermal amplification for rapid detection of Chinese sacbrood virus. J Virol Methods 176(1):115–119

    Article  CAS  Google Scholar 

  19. Li Y, Ma MX, Song Y, Li Y (2014) Development of helicase-dependent amplification method for detection of Chinese sacbrood virus. Chin J Biol 27(02):267–271

    Google Scholar 

  20. Xie Z, Qin C, Xie L, Liu J, Pang Y, Deng X et al (2010) Recombinant protein-based elisa for detection and differentiation of antibodies against avian reovirus in vaccinated and non-vaccinated chickens. J Virol Methods 165(1):108–111

    Article  CAS  Google Scholar 

  21. Chen L, Ruan F, Sun Y, Chen H (2019) Establishment of sandwich ELISA for detecting the H7 subtype influenza A virus. J Med Virol 1:25408

    Google Scholar 

  22. Zhang L, Du W, Chen C, Chen Z, Zhang L, Han Q, Xia X, Song Y, Zhang J (2018) Development and characterization of double-antibody sandwich ELISA for detection of zika virus infection. Viruses 10(11):v10110634

    Google Scholar 

Download references

Funding

This work was supported by grants from the National Natural Science Foundation of China (Nos. 31772760 and 31972626), an award for “Distinguished Professor of Liaoning Province,” the Liaoning Education Department Fund (L2015315), and the Natural Science Fund of Liaoning Province (No. 20180550289).

Author information

Author notes

  1. Ming Li and Li Sun authors contributed equally to this study.

Authors and Affiliations

  1. Jinzhou Medical University, No. 40, Section 3 Songpo Road, Jinzhou, 121001, Liaoning Province, China

    Ming Li, Li Sun, Yueyu Ma, Dongliang Fei & Mingxiao Ma

Authors
  1. Ming Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Li Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yueyu Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dongliang Fei
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mingxiao Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Dongliang Fei or Mingxiao Ma.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Handling Editor: T. K. Frey.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (xlsx 30 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, M., Sun, L., Ma, Y. et al. Development of a sandwich ELISA for the detection of Chinese sacbrood virus infection. Arch Virol 165, 1551–1556 (2020). https://doi.org/10.1007/s00705-020-04634-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-020-04634-2

Search

Navigation