TROPICSAFE partners, NIB, AGRITEST SRL and CREA, explain the basics of molecular and serological methods for detecting and characterizing phytoplasmas. Research on phytoplasmas, their biology and their epidemiology relies on our ability to detect them in plants. Since most of phytoplasmas are not yet culturable in axenic cultures, we rely on serological and molecular methods to detect and characterize them (EPPO, 2016).
The TROPICSAFE project aims to support research and diagnostics with advanced molecular and serological tools which allow pathogen specific molecular characterization. Among the promising methods for detection of phytoplasmas is LAMP (loop mediated isothermal amplification), a molecular method generating high quantities of amplified DNA from a specific DNA fragment using a set of four to six primers.
The advantages of this method include, among others, isothermal amplification which does not require expensive apparatuses, speed and high resilience to various inhibitors present in the samples. These characteristics allow fast detection of pathogens in laboratory or on-site, with minimum sample preparation. The TROPICSAFE project aims to support research and diagnostics with newly developed LAMP tests for selected pathogens. Dr. Tanja Dreo, from the TROPICSAFE slovenic partner, NIB (National Institute of Biology), share with us the work with LAMP methodlogy for detection of phytoplasmas in grapevine.
Another promising methods are ELISA and Lateral flow. The guidelines for developing diagnostic tests based on these serological methods, in terms of key issues and technical requirements, were presented by Lilia Formica and Elisa Angelini, from two of the TROPICSAFE italian partners, AGRITEST SRL and Council for Agricultural Research and Economics (CREA), respectively. The presentation covers topics concerning:
– development of antibodies against antigenic preparations of interest,
– for choosing the most suitable serological assay based on the antibody characteristics, and
– evaluation of the test quality before meeting the market.
Along the presentation, insights on the innovations exploitable for the phytoplasma project are also introduced.
You can watch these videos on the TROPICSAFE Youtube Channel!