Development of a probe-based blotting technique for the detection of tobacco streak virus.
Digoxigenin (DIG)-labeled DNA probe was developed for a sensitive and rapid detection of the tobacco streak virus (TSV) isolates in India by dot-blot and tissue print hybridization techniques. DIG-labeled DNA probe complementary to the coat protein (CP) region of TSV sunflower isolate was designed and used to detect the TSV presence at field levels. Dot-blot hybridization was used to check a large number of TSV isolates with a single probe. In addition, a sensitivity of the technique was examined with the different sample extraction methods. Another technique, the tissue blot hybridization offered a simple, reliable procedure and did not require a sample processing. Thus, both non-radioactively labeled probe techniques could facilitate the sample screening during TSV outbreaks and offer an advantage in quarantine services.
Sarovar
B
B
Department of Microbiology & Cell Biology, Indian Institute of Science, Bangalore, India. sarovarbhee@yahoo.co.in
Saigopal
D V R
DV
D036022:Capsid Proteins / Q000235:genetics; D015342:DNA Probes / Q000235:genetics*; D006368:Helianthus / Q000821:virology; D019179:Ilarvirus / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization / Q000379:methods*
D036022:Capsid Proteins; D015342:DNA Probes