A Review of Detection Methods for the Plant Viruses

Abstract

The early and accurate detection of plant viruses is an essential component to control those. Because the globalization of trade by free trade agreement (FTA) and the rapid climate change promote the country-tocountry transfer of viruses and their hosts and vectors, diagnosis of viral diseases is getting more important. Because symptoms of viral diseases are not distinct with great variety and are confused with those of abiotic stresses, symptomatic diagnosis may not be appropriate. From the last three decades, enzyme-linked immunosorbent assays (ELISAs), developed based on serological principle, have been widely used. However, ELISAs to detect plant viruses decrease due to some limitations such as availability of antibody for target virus, cost to produce antibody, requirement of large volume of sample, and time to complete ELISAs. Many advanced techniques allow overcoming demerits of ELISAs. Since the polymerase chain reaction (PCR) developed as a technique to amplify target DNA, PCR evolved to many variants with greater sensitivity than ELISAs. Many systems of plant virus detection are reviewed here, which includes immunological-based detection system, PCR techniques, and hybridization-based methods such as microarray. Some of techniques have been used in practical, while some are still under developing to get the level of confidence for actual use.

Viruses infecting common bean (Phaseolus vulgaris L.) in Tanzania: A review on molecular characterization, detection and disease management options

“…ELISA detection uses a polystyrene plate capable of binding antibodies with association of the enzymesubstrate reaction (Jeong et al, 2014). When performed using nitrocellulose and nylon membranes, it is known as a tissue blot immunoassay (Jeong et al, 2014) or dot immunobinding assay.…”

“…When performed using nitrocellulose and nylon membranes, it is known as a tissue blot immunoassay (Jeong et al, 2014) or dot immunobinding assay. With ELISA, it is possible to detect viruses in many leaf or seed samples (cost effective) in a relatively short period (normally six hours to two days).…”

Forewarned is forearmed: harmonized approaches for early detection of potentially invasive pests and pathogens in sentinel plantings

“…Detection and identification of phytoplasma and spiroplasma are primarily based on 16S rRNA (16Sr) amplification followed by restriction fragment length polymorphism analysis (Bertaccini et al 2019). When genetic information is available, PCR and reverse transcription PCR are used to detect plant viruses (Jeong et al 2014).…”

Coinfección natural de virus de ARN en cultivos de papa (<i>Solanum tuberosum</i> subsp. Andigena) en Antioquia (Colombia)

“…En la actualidad, la prueba de ELISA se considera el test Gold Standard en los procesos de certificación de tubérculossemilla de papa y para el diagnóstico de virus de importancia cuarentenaria (Halterman et al, 2012;Frost et al, 2013;Stammler et al, 2018); sin embargo, en este estudio se encontró que la técnica de ELISA detectó un menor número de muestras infectadas con los virus PVY (55,5 %), PLRV (5,5 %) y PVX (0 %). Estos resultados no resultan sorpresivos, por cuanto existen reportes que señalan que la sensibilidad de la RT-qPCR para la detección de virus de plantas varía entre 10.000 y 100.000.000 veces más que la prueba de ELISA (Agindotan et al, 2007;Joo-jin et al, 2014;Nikitin et al, 2018;Stammler et al, 2018) y dicha observación se ha confirmado también en diferentes trabajos realizados en Antioquia sobre virus de papa. Así por ejemplo, García Ruíz et al, (2016) reportaron niveles de infección con el virus PVX en un 93,75 % de muestras de tubérculos-semilla de papa y papa criolla cuando emplearon RT-qPCR, mientras que al usar la técnica de ELISA, sólo detectaron el virus en el 50 % de éstas.…”