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Turkish Journal of Biology

Abstract

Background/Aim: Sugarcane mosaic virus (SCMV; genus Potyvirus and family Potyviridae), poses a significant threat to global sugarcane cultivars, including those in Pakistan. The aim of this study was to develop a rapid and effective diagnostic tool for detection of SCMV, enabling timely implementation of control measures to mitigate potential yield losses.Materials and Methods: The study focused on the in silico analysis; physicochemical properties, immunogenicity and subcellular localization of the SCMV coat protein (CP). The SCMV coat protein (CP) gene was synthesized, cloned and expressed in Escherichia coli. The recombinant fusion coat protein (rFCP-SCMV) was purified and used to generate polyclonal antibodies (pAB) in mice. The immunogenicity of the antibodies was evaluated through indirect ELISA and RT-PCR.Results: Epitope prediction using tools like the OptimumAntigen™ design tool from GenScript, BepiPred and IEDB identified key B-cell epitopes on the SCMV coat protein, enhancing the specificity of the antibodies. Structural modeling with SWISS-MODEL and PyMOL provided insights into the 3D structures of viral proteins and their epitopes, aiding in the design of high-affinity antibodies. Molecular docking studies simulated the interaction between antibodies and viral epitopes, allowing for the selection of optimal antibody candidates. The synthesized recombinant fusion coat protein (rFCP-SCMV) was used to produce polyclonal antibodies (pAb) in mice. These antibodies exhibited high sensitivity, detecting as low as 100 pg of SCMV protein in indirect ELISA assays. They also effectively identified SCMV in infected sugarcane field samples, confirmed by RT-PCR. The antibodies maintained high specificity and sensitivity even at a 1:10,000 dilution, proving their efficacy in recognizing both the recombinant protein and virus particles in plant sap.Conclusion: The study reveals a rapid, effective immunodiagnostic technique for detecting SCMV in sugarcane cultivars, offering an accurate alternative to conventional virology methods, reducing contamination risk, and providing a valuable tool for mitigating yield losses.

Author ORCID Identifier

HAFIZ KHALID: 0009-0000-8407-0660

NAJAM US SAHAR SADAF ZAIDI: 0009-0008-1940-5100

NAEEM RASHID: 0000-0002-3071-5977

MUHAMMAD TAHIR: 0000-0002-4025-852X

DOI

10.55730/1300-0152.2714

Keywords

Sugarcane mosaic virus, polyclonal antibody, RT-PCR, immunodiagnostic, sugarcane

First Page

390

Last Page

400

Creative Commons License

Creative Commons Attribution 4.0 International License
This work is licensed under a Creative Commons Attribution 4.0 International License.

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