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Comparison of DAC-ELISA and duplex IC-RT-PCR for the diagnosis of Sugarcane streak mosaic virus and Sugarcane mosaic virus in sugarcane

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Title Comparison of DAC-ELISA and duplex IC-RT-PCR for the diagnosis of Sugarcane streak mosaic virus and Sugarcane mosaic virus in sugarcane
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Creator Scindiya, M., V. Ganesh Kumar, R. Karuppaiah and R. Viswanathan
 
Subject IC-RT-PCR, Sugarcane Streak Mosaic Virus (SCSMV), DAC-ELISA,
 
Description Not Available
Studies were conducted to standardize IC-RT-PCR to detect SCSMV and SCMV with more specificity and sensitivity. A total of 30 samples of different varieties of sugarcane which varied in symptoms from severe mosaic symptoms to asymptomatic were diagnosed by DAC-ELISA using recombinant SCSMV coat protein antiserum in which it gave both positive and negative values. Some of the samples with mild symptoms were found to be negative in the assay, hence it warranted further confirmation of negative samples employing molecular tools. In order to accomplish this, a duplex IC-RT-PCR was developed for a simultaneous diagnosis of both the viruses in a single reaction. From DAC-ELISA results, 12 negative samples were diagnosed by duplex IC-RT-PCR using recombinant SCSMV-coat protein antiserum. Among them, eight samples were positive to SCSMV and eleven samples were positive to SCMV, when diagnosed by IC-RT-PCR which indicated that this technique is more sensitive than DAC-ELISA and the results are illustrated by specific bands to SCSMV (~690bp) and SCMV (~380bp) in a single reaction. The antiserum was found to trap both the viruses in IC-RT-PCR. Our assays very clearly indicated that five varieties were positive to SCSMV in both DAC-ELISA and IC-RT-PCR, which confirms that these samples are infected only by SCSMV and not by SCMV. Another four varieties showed mixed infections of the two viruses when diagnosed by duplex IC-RT-PCR. Comparison of IC-RT-PCR with DAC-ELISA revealed that the samples which are negative in DAC-ELISA were positive in IC-RT-PCR and that establishes higher sensitivity of the former technique. Although DAC-ELISA was found to be simple and cost effective to diagnose the virus, in samples with very low titre, IC-RT-PCR would be a more sensitive technique to detect the virus(es). Our study very clearly establishes that IC-RT-PCR is more sensitive to detect the viruses present in low titre more specifically than DAC-ELISA.
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Date 2021-08-26T05:53:59Z
2021-08-26T05:53:59Z
2012-10-15
 
Type Presentation
 
Identifier Not Available
Not Available
International Symposium on New Paradigms in Sugarcane Research
http://krishi.icar.gov.in/jspui/handle/123456789/60607
 
Language English
 
Relation Not Available;
 
Publisher Society for Sugarcane Research and Development, Coimbatore, India