Development of recombinant coat protein antibody based IC-RT-PCR for detection and discrimination of sugarcane streak mosaic virus isolates from Southern India.
Sugarcane streak mosaic virus (SCSMV), causes mosaic disease of sugarcane and is thought to belong to a new undescribed genus in the family Potyviridae. The coat protein (CP) gene from the Andhra Pradesh (AP) isolate of SCSMV (SCSMV-AP) was cloned and expressed in Escherichia coli. The recombinant coat protein was used to raise high quality antiserum. The CP antiserum was used to develop an immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) based assay for the detection and discrimination of SCSMV isolates in South India. The sequence of the cloned PCR products encoding 3'untranslated region (UTR) and CP regions of the virus isolates from three different locations in South India viz. Tanuku (Coastal Andhra Pradesh), Coimbatore (Tamil Nadu) and Hospet (Karnataka) was compared with that of SCSMV-AP. The analysis showed that they share 89.4, 89.5 and 90% identity respectively at the nucleotide level. This suggests that the isolates causing mosaic disease of sugarcane in South India are indeed strains of SCSMV. In addition, the sensitivity of the IC-RT-PCR was compared with direct antigen coating-enzyme linked immunosorbent assay (DAC-ELISA) and dot-blot immunobinding assays and was found to be more sensitive and hence could be used to detect the presence of virus in sugarcane breeding, germplasm centres and in quarantine programs.
Hema
M
M
Department of Virology, Sri Venkateswara University, Tirupati, Andhra Pradesh, India.
Kirthi
N
N
Sreenivasulu
P
P
Savithri
H S
HS
D000914:Antibodies, Viral / Q000276:immunology*; D036022:Capsid Proteins / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D015151:Immunoblotting; D007194:India; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000821:virology; D010942:Plant Viruses / Q000276:immunology* / Q000302:isolation & purification; D019176:Potyviridae / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D031786:Saccharum / Q000821:virology*; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D036022:Capsid Proteins