LAMP, or Loop Mediated Isothermal Amplification is a DNA fingerprinting technology which, unlike qPCR, it doesn’t require thermocycling to amplify the target DNA . This has allowed development of simple, portable, low cost devices for rapid bioanalysis in field environments. FERA has been working with key collaborators on an Innovate UK project , evaluating the Genie III portable LAMP platform to provide a DNA diagnosis of Septoria tritici in crops. They have also looked at bacterial rots in strawberries and peas.
(NOTE: Similar portable technology for qPCR has also recently been developed by Biomeme, but not yet tested for arable crop pathogens. The next generation of portable PCR devices use digital PCR (dPCR) technology, and researchers at Zhejiung University are developing portable quantitative devices using smartphone connectivity).
How it Works
Leaf samples are mechanically macerated and introduced into the machine. Reagents using 4 different primers recognising 6 regions on the target gene, DNA polymerase with strand displacement activity, and all necessary reaction substrates are all included in a single mixture. The reaction occurs at a constant 65°C using a strand displacement reaction, a detectable by-product of which is produced in proportion to the amplified DNA and can be semi-quantified.
Farmer and Agronomist Benefits
Systems are quick (30mins) inexpensive and easy-to-use, allowing real-time monitoring of crop diseases and their fungicide resistance status. This supports a more targeted approach to timing and selection of fungicide applications, which optimises fungicide efficacy and reduces resistance build-up. Growers save money on ineffective sprays and increase yield and quality alongside sustainable production.