title: Immunodiagnosis of episomal Banana streak MY virus using polyclonal antibodies to an expressed putative coat protein.
abstract: A cryptic Badnavirus species complex, known as banana streak viruses (BSV) poses a serious threat to banana production and genetic improvement worldwide. Due to the presence of integrated BSV sequences in the banana genome, routine detection is largely based on serological and nucleo-serological diagnostic methods which require high titre specific polyclonal antiserum. Viral structural proteins like coat protein (CP) are the best target for in vitro expression, to be used as antigen for antiserum production. However, in badnaviruses precise CP sequences are not known. In this study, two putative CP coding regions (p48 and p37) of Banana streak MY virus (BSMYV) were identified in silico by comparison with caulimoviruses, retroviruses and Rice tungro bacilliform virus. The putative CP coding region (p37) was in vitro expressed in pMAL system and affinity purified. The purified fusion protein was used as antigen for raising polyclonal antiserum in rabbit. The specificity of antiserum was confirmed in Western blots, immunosorbent electron microscopy (ISEM) and antigen coated plate-enzyme linked immunosorbent assay (ACP-ELISA). The antiserum (1:2000) was successfully used in ACP-ELISA for specific detection of BSMYV infection in field and tissue culture raised banana plants. The antiserum was also utilized in immuno-capture PCR (IC-PCR) based indexing of episomal BSMYV infection. This is the first report of in silico identification of putative CP region of BSMYV, production of polyclonal antiserum against recombinant p37 and its successful use in immunodetection.
authors: Advanced Centre for Plant Virology, Division of Plant Pathology, Indian Agricultural Research Institute, Pusa, New Delhi 110012, India.
authors: Susheel Kumar
authors: SK
authors: Sharma
authors: Advanced Centre for Plant Virology, Division of Plant Pathology, Indian Agricultural Research Institute, Pusa, New Delhi 110012, India.
authors: P Vignesh
authors: PV
authors: Kumar
authors: Advanced Centre for Plant Virology, Division of Plant Pathology, Indian Agricultural Research Institute, Pusa, New Delhi 110012, India. Electronic address: vbaranwal2001@yahoo.com.
authors: Virendra Kumar
authors: VK
authors: Baranwal
mesh_terms: D000818:Animals; D000914:Antibodies, Viral; D019195:Badnavirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology*; D019295:Computational Biology; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D007159:Immunologic Tests / Q000379:methods; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D028521:Musa / Q000821:virology*; D016366:Open Reading Frames; D010935:Plant Diseases / Q000821:virology*; D011817:Rabbits; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins
keywords: ACP–ELISA; Banana streak MY virus; IC-PCR; Indexing; Polyclonal antibodies; Putative coat protein