Comparative analysis of ELISA, one step RT-PCR and IC-RT-PCR for the detection of bean yellow Mosaic virus in gladiolus.
To overcome the problematic detection of Bean yellow mosaic virus (BYMV) concentration in gladiolus corms or cormlets, we tested suitability of different diagnostic methods such as ELISA, one step RT-PCR and IC-RT-PCR. Among these methods, DAS-ELISA and one step RT-PCR method was able to detect virus in leaves and it failed to detect in corms or cormlets whereas IC-RT-PCR method was very sensitive and able to detect virus in both leaves and corms or cormlets of gladiolus plants this is due to during an RNA isolation step and viral particles are enriched by antibody or plastic affinity capture whilst PCR inhibitors are eliminated from the sample during the immunocpature stage of the test. Therefore, IC-RT-PCR was the most reliable method of diagnosing BYMV in gladiolus corms or cormlets.
Ganesh Selvaraj
Duraisamy
D
Mendel University of Agriculture and Forestry, Faculty of Agronomy Department of Crop Science, Breeding and Plant Medicine Zemĕdĕlská 1, CZ-613 00 Brno, Czech Republic. snow123@ymail.com
Pokorný
Radován
R
Holková
Ludmila
L
D018556:Crops, Agricultural / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D035264:Flowers / Q000821:virology*; D005784:Gene Amplification; D005814:Genes, Viral / Q000235:genetics; D009029:Mosaic Viruses / Q000235:genetics* / Q000302:isolation & purification*; D018515:Plant Leaves / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods