LAMP for detection of ToBRFV (PE 035)

ToBRFV is a new virus representing a continuing emergency threat to UK tomato production (value £200m; TGA retail estimate) and is a notifiable disease requiring statutory action if outbreaks occur. Currently, surveillance for this virus involves sampling large numbers of plants from a glasshouse which is time consuming for the Plant Health Inspector, costly, risks further unnecessary spread should an outbreak occur, and only gives a ‘snapshot’ of a single point in the season. An alternate approach, such as ongoing monitoring of critical points in the process, using LAMP (Loop-mediated isothermal amplification) diagnostics for testing swabs and irrigation water, would potentially allow a more comprehensive monitoring approach to be followed throughout the season for less cost. Additionally, current AHDB funded research on detection of the virus in planta (PE 034) is showing that infections entering the plant later in the growing season may not be reliably detected, supporting the need for alternative diagnostic approaches for glasshouse and packhouse monitoring.  

Defra are currently funding a project under the “Future Proofing Plant Health” framework to validate LAMP PCR diagnostics for “non-invasive” onsite detection of ToBRFV. This Defra funding is primarily aimed at validation for use by inspection services, however, the same technology and approaches could be used by growers for ongoing monitoring for this pathogen.  This project will move the technology to a near market position for this application through a pilot study with a limited number of growers, and the technology company (Optigene) will be engaged to pick up developments for use by growers. An additional benefit of this work will be that the project should provide a framework for future diagnostic developments to benefit both tomato growers and the wider protected edibles industry through an increased range of diagnostic targets and potential for moving the technology into a format for “in-line” testing in production systems. 

The aim of the project is to demonstrate the value of environmental sampling (“non-invasive sampling” or “non-plant sampling”) for on-site monitoring of plant health status of crops with respect to ToBRFV infection.  This will be achieved through the following objectives:  

1) Validation of ToBRFV LAMP for onsite testing, including trial “process control” for presence of pepino mosaic virus (PepMV).  

2) Test industry and “mock” glasshouse samples submitted to Fera to beta-test the processing and testing approaches  

3) Produce user guidance covering sampling and testing of samples for on-site testing.  

4) Stakeholder workshops to demonstrate testing platform.