Proposed Implementation
Since the beginning of our project we envisioned Diagnosgene as a point-of-care detection system for easy execution on the field. For our proposed implementation, the target users are farmers in the local communities of Mexico. Agave is produced in over 550 municipalities by Mexican farmers in need of this new strategy proposed.
We envision our project being an easy-to-use, portable, accessible and quick-results diagnostic product for Fusarium oxysporum that can be used by the civil associations and farmers. Diagnosgene allows farmers and anybody to perform the tests themselves and have a result in less than 3 hours without the need of specialized equipment or personnel. We suggest that the agave farmers use our product when they buy a young plant (pups) in order for them to be sure that it is not sick before they plant it in their land and it contaminates others. You can check out our Human Practices to read about the different ways the farmers can acquire their agave pups. We also think it best for the farmers to perform sporadic tests with our product throughout the growth of the plants to ensure there is no infection.
We also have to take into account the data described in our Human Practices page, which includes the findings of an unfair treatment from big Tequila companies towards agave farmers, where the payment at the time of harvesting the plants can be much less than 10% of the total price of the agave without any consequences. This means that while distributing our product we should assure collaborations with socially responsible companies that are transparent on their actions and fair work policies. Our intended entrepreneurship for implementing our project consists of a Business to Business model that the agave producer at the center (see more on Entrepreneurship).
Once we finish with the laboratory experimentation that is left, we plan on developing a prototype of our detection system that includes a DNA extraction device and the necessary components to perform both the RPA amplification and the cell-free reaction (see more on Design). Once we validate our prototype with producers we are planning on selling our final product as a commercial detection kit with all the necessary materials and reagents as well as with a comprehensive user manual that facilitates the use for farmers.
As for safety aspects to take into consideration, instead of using a living organism as a biosensor, we decided to go for a cell-free system, which is safer to use on the field and for people not specialized on the subject, this way there can not be any GMO spread. However to develop the cell-free a chassis is needed, so we are using an organism on a biosafety level 1 (E. coli BL21). You can check out more about it on our Design page, and read about our laboratory safety at the Safety page. As for our proof of concept, we decided that using a short non-coding sequence from Agave tequilana and Fusarium oxysporum to test the toehold switches was the best option, instead of testing them on the actual infected plant, which could cause the fungi to spread and be harmful. For our human practices, we did not use any names without authorization and gave anonymous identities to the agave farmers who helped us.
