Antibody and nucleic acid probe-based techniques for detection of sugarcane streak mosaic virus causing mosaic disease of sugarcane in India

Hema, M and Savithri, HS and Sreenivasulu, P (2001) Antibody and nucleic acid probe-based techniques for detection of sugarcane streak mosaic virus causing mosaic disease of sugarcane in India. In: Current Science, 81 (8). pp. 1105-1108.

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Abstract

Double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) and direct antigen coating (DAC)-ELISA tests were evaluated for detection of sugarcane streak mosaic virus (SCSMV-AP), a new member of Tritimovirus genus in the family Potyviridae, in leaf extracts, sugarcane juice and purified virus. The virus was detected up to 1/3125 and 1/625 dilutions in infected sugarcane leaf, 5 mul and 10 mul/well in sugarcane juice, 1/3125 and 1/3125 dilutions in infected sorghum leaf and 10 ng and 50 ng/ml for purified virus in DAS-ELISA and DAC- ELISA tests, respectively. A cDNA clone pSCSMV-AP (495 bp) specific to SCSMV-AP was selected as diagnostic P-32 and DIG (digoxigenin) probe. In slot-blot hybridization analysis, P-32 and DIG probes reacted with total nucleic acid extracts from infected sugarcane leaf (10(-5) and 10(-4)), infected sugarcane juice (10(-3) and 10(-4)) and infected sorghum leaf (10(-5) and 10(-5)). With both the probes the virus was detected in purified virus up to 50 pg and viral RNA up to 1 pg level. DAS- ELISA appears to be sensitive and ideal for routine large-scale detection of virus in small leaf tissue and cane juice samples of sugarcane. Nucleic acid-based tests could be useful in screening of sugarcane germplasm.

Item Type:	Journal Article
Publication:	Current Science
Publisher:	Current Science Assn
Additional Information:	Copyright for this article belongs to Indian Academy of Sciences
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	15 Sep 2004
Last Modified:	19 Sep 2010 04:16
URI:	http://eprints.iisc.ac.in/id/eprint/1914

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