Non‐radioactive molecular hybridization detection of carnation mottle virus in infected carnations and its comparison to serological and biological techniques

Abstract

A non-radioactive molecular hybridization method was developed to detect carnation mottle virus (CarMV) in carnation plants. This method was compared to the standard ELISA test and biological assays. For crude plant extracts, the molecular hybridization technique proved to be at least 125 times more sensitive than double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). The method was adapted to avoid the necessity to clarify the samples by centrifugation and to use pipettes to load them. By introducing this modification, the hybridization assay was found to be a suitable method for routine virus detection. The relative benefits of using this procedure for large-scale indexing are discussed.

Plant tissue distribution and chemical inactivation of six carnation viruses

“…Detection at early stages of viral infection and subsequent elimination of the infected material constitute the main approach to combating those disorders. In this sense, incorporation of detection techniques, such as nonradioactive molecular hybridisation , into the routine diagnosis of carnation crops has increased the detection limit usually obtained with widely used serological assays (Sa´nchez-Navarro et al, 1996. However, studies addressed to elucidate the viral distribution in the infected plant or to inactivate the virus with different chemicals are sparse.…”

Biological, Serological and Molecular Characterization of Papper MildMottle Virus Isolated from Weast Region of Kingdom of Saudi Arabia

“…These methods have replaced the traditional methods based on biological properties. PCR assays using certain primers presented reliable and sensitive means for detection and characterization of the plant pathogenic [6,49].…”

Occurrence and Geographical Distribution of the ‘Torrado’ Disease in Spain

“…Given the high percentage of ToTV‐infected tomato plants and the lack of an available commercial antibody, we explored alternative detection techniques to the standard serological ELISA method to be used to screen a large amount of samples. Molecular hybridization (MH) using non‐radioactive riboprobes has proved to be an interesting detection technique for carnation (Sánchez‐Navarro et al. 1996), stone fruit (Herranz et al.…”

“…Given the high percentage of ToTV-infected tomato plants and the lack of an available commercial antibody, we explored alternative detection techniques to the standard serological ELISA method to be used to screen a large amount of samples. Molecular hybridization (MH) using non-radioactive riboprobes has proved to be an interesting detection technique for carnation (Sa´nchez-Navarro et al 1996), stone fruit ) and tomato crops (Saldarelli et al 1996). The analysis of the five ToTV Spanish isolates reveals that the fragment which includes the putative MP in the ORF2-RNA2, is a conserved region that represents a good viral portion to be detected by MH.…”