Recombinant of Sugarcane streak mosaic virus Coat Protein Polyclonal Antiserum Production and Its Serological Assays: Produksi Poliklonal Antiserum Rekombinan Protein Selubung Sugarcane streak mosaic virus dan Uji Serologinya

Hamdayanty; Sri Hendrastuti  Hidayat; Tri Asmira Damayanti

doi:10.14692/jfi.22.1.31-39

Authors

Hamdayanty Department of Plant Protection, Faculty of Agriculture, IPB University
Sri Hendrastuti Hidayat Department of Plant Protection, Faculty of Agriculture, IPB University
Tri Asmira Damayanti Departemen Proteksi Tanaman, Fakultas Pertanian, IPB

DOI:

https://doi.org/10.14692/jfi.22.1.31-39

Keywords:

antiserum, protein, recombinant CP gene, streak mosaic, sugarcane

Abstract

Sugarcane streak mosaic virus (SCSMV) is one of the important viruses that causes streak mosaic disease in sugarcane plants. This virus spreads rapidly mechanically and is transmitted by cutting canes. Until now, a commercial SCSMV antiserum has not been available, even though it is essential for routine field detection and monitoring of disease development. A recombinant clone of the SCSMV coat protein gene (SCSMV-CP) in an expression vector has previously been successfully synthesized. This study aimed to produce a polyclonal antiserum by overexpressing recombinant SCSMV-CP in the Escherichia coli strain BL21(DE3) as an antigen and to test the antiserum’s sensitivity for detecting SCSMV serologically. The sensitivity and specificity of the recombinant antiserum for detecting SCSMV were determined using the agar gel precipitation test (AGPT), Indirect ELISA (I-ELISA), and dot blot immunobinding assay (DBIA) methods. Recombinant SCSMV-CP antiserum was successfully produced in New Zealand rabbits, but it failed to detect SCSMV using the AGPT method. The sensitivity of SCSMV antiserum in the I-ELISA method was best up to an antiserum dilution of 1:1000 (v/v), and the sensitivity of the DBIA method was higher up to an antiserum dilution of 1:10 000 (v/v). SCSMV antiserum reacted negatively to SCMV antigen in the I-ELISA method, indicating the high specificity of the recombinant SCSMV-CP antiserum. This recombinant antiserum is suitable for routine detection of SCSMV using both I-ELISA and DBIA methods and has the potential for future mass production. The DBIA detection method is suitable for large-scale samples, is easy, fast, and accurate, and is cheaper than I-ELISA.

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