Going beyond Ideas

Design & Building

Designing a test kit that can be used outside the lab can be an arduous task and one should always tread carefully. Various chemical substances have the caveat that they cannot be used without proper equipment or prior expertise. Moreover chemicals that are harmful to the environment or the user must be excluded. Additionally, using low cost substances and techniques in the test kit is of primordial importance to us since it constitutes one of the most important advantages in comparison with existing detection techniques. Last but not least, all the reagents used must be able to sustain long-distance transportation so as not to arrive in foul condition in their end destination.

For that reason we sought to pinpoint the best reagents and tools that would be required in our kit individually.

After careful consideration, conversations with agricultural scientists, literature search and small experiments, that you can find below, we decided that each kit would include the following:

3 Eppendorf tubes(1.5ml) each containing:

  • 0.3 ml of the lysis buffer (50% ethanol/2M NaCl) with a modified cap for easy content discharge
  • 0.7 ml of our fixated engineered e-coli cells
  • 1 ml of ethanol 100% sealed hermetically

3 Eppendorf tubes(1.5ml) each containing:

1 test strip attached to a 3D PLA printed platform in the same fashion as the one found in SARS-CoV-2 tests

1 Instruction manual

In order to find the best possible lysis buffer that would simultaneously be cheap to produce and safe for users we devised the following experiments:

After a thorough search through literature during which we aimed to find the quickest and highest yielding method for plant cell lysis1,2, we narrowed down our results to the ones that needed no special equipment or temperatures above or below room temperature. Moreover we tried to use variations of known cell lysis buffers as standalone buffers that would normally be part of a greater protocol.

After a thorough search through literature during which we aimed to find the quickest and highest yielding method for plant cell lysis 1,2, we narrowed down our results to the ones that needed no special equipment or temperatures above or below room temperature. Moreover we tried to use variations of known cell lysis buffers as standalone buffers that would normally be part of a greater protocol.

Leaves from a single Arabidopsis thaliana plant were collected and cut down in approximately equal pieces (Image 1)

For each solution tested two methods of mechanical breaking down was used: one with a plastic pestle and one with a toothpick (Image 2).

After 10 mins have passed, in each buffer containing the lysed cells, a Trypan blue 4% solution was added to stain dead cells. 25 ul of each sample was then immediately placed on a Neubauer slide and the ratios of dead to total cells were counted (Image 3).

Results are presented below:

Solution Type of tool used D/total cells Ratio Weighted D/total cells aka ratio-contribution of solution
NaOH 0.1M/SDS 2% Pestle 0.535433071 1.255498235
NaOH 0.1M/SDS 2% Toothpick 0.568181818 3.357438017
EDTA 20 uM/3.5M NaCl Pestle 0.607594937 1.424705369
EDTA 20 uM/3.5M NaCl Toothpick 0.210084034 1.241405653
EDTA 20 uM/1.4M NaCl Pestle 0.222222222 0.521072797
EDTA 20 uM/1.4M NaCl Toothpick 0.254545455 1.504132231
EtOH 50%/2M NaCl Pestle 0.73255814 1.717722534
EtOH 50%/ 2M NaCl Toothpick 0.621621622 3.673218673
EtOH 70%/2M NaCl Pestle 0.702290076 1.646749145
EtOH 70%/2M NaCl Toothpick 0.348837209 2.061310782
NaCl 2M Pestle 0.513513514 1.204100652
NaCl 2M Toothpick 0.320895522 1.896200814
Positive Control (freeze thaw)   0.863636364  
Negative Control (dH20) Pestle 0.426470588  
Negative Control (dH20) Toothpick 0.169230769  

With a quick look over the data one can easily infer that the plastic pestle is by far superior to a toothpick even in dH20 alone. Thus, given that a pestle will be used, the best solution option is EtOH 50%/2M NaCl.

User Manual

Lastly, we designed the instruction manual that includes all needed information in a simple yet precise manner for anyone to use. We aimed to streamline the process by naming all solutions used as S1,S2 and S3 and we also included a key for the interpretation of results. It is our belief that we simplified the process as best as we could without sacrificing any sort of scientific validity.

Identifying global needs


As with every virus, TSWV has its own distinct epidemiology, affecting some countries more than others. That is the reason that we sought to pinpoint the countries most heavily affected by the pathogen. After a thorough search we stumbled across some very well upkept databases that included data on the spread of tospoviruses around the world as well as their future projected status. A great model country to study the epidemiology of TSWV would be Egypt since it occupies a large portion of the worldwide tomato production but data, compared to other countries, are not easily retrievable.

According to FAO and Hagar Saeed, journalist of the “Egyptian Gazette”, the area of agricultural land in Egypt is confined to the Nile Valley and delta, with a few oases and some arable land in Sinai. The total cultivated area estimations range from 7.2 to 11.5 million feddans (1 feddan = 0.42 ha), depending on the year and the definition of what constitutes a cultivated land. Agriculture is a major component of the Egyptian economy, contributing 11.3 % of the country’s gross domestic product. The agricultural sector accounts for 28 % of all jobs and over 55 % of employment in Upper Egypt is agriculture-related.

It is important to note that the extent of the damage that solely TSWV has caused is unknown, yet it surely contributes to the plummet observed.

Nonetheless, one should always keep in mind that TSWV also affects pepper, lettuce, potato, papaya, groundnut, tobacco and chrysanthemum crops but most importantly, TSWV has an extremely wide host range with more than 1300 wild and weed species5,6. This includes plant species in 15 monocotyledonous and 69 dicotyledonous families and one family of the Pteridophyta making it a major danger in our efforts for environmental conservation. Given that a plant can be a TSWV host even post-harvest makes it extremely easy to spread as the following map indicates

It is our personal opinion that even some countries not highlighted are affected by the virus yet either the presence of the virus has gone undetected or the spread has happened recently and data is still not available.

In any case there is no doubt that this virus is a global concern that, for now, affects some countries more.


With an ideal candidate country in mind (Egypt) we sought out to identify the best possible way of distribution to it and what would that hypothetical deployment would require.

Distribution methods are heavily limited by the fact that the test includes live cells. Even though the cells, per se, are fixated and thus minimal danger is posed in regards to their survivability, the fact that they remain alive is enough of a liability for transportation standards. This is why we sought to learn how live material is preserved and transported in countries like Egypt and China. After short communications with people on the transportation sector we were informed that each country has its own distinct laws when it comes to importing and/or quarantining live material from other countries. We decided that it would be more realistic to focus our efforts in our neighbouring country of Egypt since actual transportation of our manufactured test there would be more realistic.

After reaching out to transportation companies and legal advisors we came to the conclusion that it would be wiser to ship our tests in bulk. Each test kit would be put in a hermetically sealed envelope and all envelopes would be placed in a 20*20*20 centimetre box. Each box would fit 15 kits and the average cost of transportation after enquiring about the cost in 8 different transportation companies was calculated to be 65.45 € thus amounting to a transportation cost of 4.36 € per kit. In addition to the aforementioned cost of production of the kit the grand total for the production and delivering of the test is 5.46 € .

Up next we decided to investigate the intricacies involved with transporting genetically modified microorganisms (GMMOs or GMMs). Depending on the mode of transportation, the country of origin and arrival, laws may vary. In our case we had to cover each part separately.

Departing from Greece

The laws of Greece regarding the export of GMMOs have been replaced and are in total alignment with the ones proposed by EU. More specifically according to “Directive 2001/18”, “Regulation 1829/2003”, “Regulation 1830/2003” and “Directive 90/219/EEC” even though the deployment of such organisms in EU soil is strictly regulated and a final decision on its usage may take up to 10 years. However, regarding transportation laws regulate that a legal entity is to be always held accountable for the transportation after a thorough quality check and an approval system that is yet to be deployed in its final form. After consulting with various experts on the field we have been informed that such an approval process may take up to a few years, giving us enough time to perfect our project and minimise any risk.


Rules regarding the transportation of GMMOs are set by the IATA Dangerous Goods Regulations (62nd Edition) that is effective since the 1st of January 2021. Genetically modified organisms and microorganisms which do not meet the definition of toxic or infectious substances (such as ours is) must be assigned to code UN 3245. Boxes and individual contents must be shipped following Packing Instruction P904 (ICAO/IATA PI959) with a proper shipping name. This can be UN 3245 followed either by “GENETICALLY MODIFIED MICRO- ORGANISMS” or "GENETICALLY MODIFIED ORGANISMS".

Arriving in Egypt

According to the Library of Congress of the U.S “Egypt has no restrictions on releasing genetically modified organisms into the environment. In March 2008, the Ministry of Agriculture approved the domestic cultivation of genetically modified corn, and the Egyptian Ministry of Agriculture allowed the importation of twenty-eight tons of genetically modified corn seeds into Egyptian markets. However, in the spring of 2009, genetically modified corn seed imports were halted so that the National Biosafety Committee (NBC) could complete the country’s National Biosafety Framework (though the NBC continued to permit the planting of locally produced biotech seeds in newly reclaimed areas)”. Moreover, via our own research we could not find any special laws that have been drafted and voted by local or national assemblies on the importation and deployment of GMMOs.

In conclusion it is our firm belief that realistically speaking, the most austere measures that need to be abided to are the ones set by the E.U, yet the lack of appropriate laws by the Egyptian government must not take away from the serious nature with which we need to design our test kit in order to avoid any accidental deployment of our living yet fixated genetically modified e-coli cells in Egyptian soil.

Once all requirements are met it would be wise to look to more countries on their respective laws. However, it is a good practice to always abide to the strictest of laws regarding GMMOs and build the detection kit accordingly in order to avoid future obs.

Scaling Up

Italy presents itself as a prime candidate for Mediterranean and Alpine biodiversity with rapid geological switches between ecosystems. Due to its prime location in the centre of Southern Europe, its proximity to Greece and the fact that it is a member of the European Union, Italy is an ideal place for setting up our first outsourced European taskforce.

This part of the implementation would be the most primordial if our project was purely diagnostic. Nonetheless, given that our team, in the end of the day, proposes a general approach in conservation strategies of the future renders this part of our implementation strategy just as important as any other part. With no errors in judgement one can easily infer the fact that implementation for one type of plant, either crop or wild, equals implementation for all. This fact stems from the “one-test-fits-all” character of our project as well as the relatively uniform nature of laws regarding its usage. Ergo, the following hypothetical 10-year plan was drafted by our team concerning the gradual establishment of our tool as a way to track and aptly face plant epidemics.

Years 1-3

Actions of the team throughout the first years would not look much different than the ones performed at the moment. In a larger timeframe our team would be able to work constantly on troubleshooting any major or minor difficulties our kit would face in the real world while searching for the additional funding that would be required to carry them out. Moreover throughout these years, through conversations with law experts, EU lawmakers and relevant independent think tanks and groups we would be able to draft out and commence all the required processes so we abide by, at least, EU law by the time of the first field usage.

In the meantime, our R&D team would pinpoint all the available plant species or their crop relatives that are impacted by TSWV and draft multiple comparative tests in an enclosed lab environment in order to quantify our kit’s efficacy as a whole. Additional methods to streamline the tool’s production would be investigated as well.

Large Scale

Year 4-6

Given that all open-ends of our project have been located and resolved, in these years our team would conduct the set of experiments proposed in the following table. Please note that some experiments such as 2 and 3 will coincide for some of their duration. Please advise the timeline provided after the table for full information.

Experiment No Scale Plant species Location Length User
1 Small 1 Crop fields 3 months Our team
2 Small 1 Crop fields 6 months Certified Personnel
3 Medium 2-10 Crop fields 1 year Farmers
4 Small 1 Wild 6 months Our team
5 Large 10-30 Crop fields 15 months Farmers
6 Medium 2-10 Wild 2 months Our team
7 Medium 2-10 Wild 1 year Certified Personnel

In the middle of the period a gradual shift can be observed towards the wild. This shift is completely intentional and ties entirely with our plans for years 7-10.

Final Goal

Years 7-10

As we have already stated, our final goal is to provide a viable solution for conservation efforts around the globe. After successfully completing the aforementioned experiments our team would find itself in an advantageous position; by having established our tool in the field of agrodiagnostics our team would have the necessary funds to redirect towards conservation efforts. By using the income generated from crop conservation efforts we can:

1.   Conduct the experiments needed for the repurposing of our kit to diagnose even more phytopathogens

2.   Establish permanent task force teams in places of interest for reconnaissance expeditions in places of rich wilderness

1. Repurposing

Even though TSWV, and tospoviruses in general, certainly pose a great threat to biodiversity, a plethora of phytopathogens exist that have the potential to harm wild plants or they already have. Microorganisms such as Phytopthora infestans, Ips typographus, Anoplophora chinensis, Thaumetopoea pityocampa and many more are becoming an increasing threat in forests, especially the ones in the Holarktis region. According to the European Commission’s (DG ENV) final report on the disturbances of EU forests caused by biotic agents (2012)3 these pathogens exhibit an aggressive spread and demonstrate a large spectrum of minima and maxima of survivability. Accordingly in Neotropis and Paleotropis, several dangerous pathogenic species have been identified. Notably in the Amazon Forest, certain soils reveal susceptibility to phytopathogens and lower fungal community dissimilarity than other forests4 thus making it a prime candidate for the proliferation of harmful microbial species.

Repurposing the detection kit won’t be an easy task. Nonetheless, given that our R&D team will only have this single task as their only preoccupation, combined with their projected gained expertise throughout the past 7 years, a proper full-out repurposing seems realistic.

2. Taskforces

Without a doubt, phytopathogens infiltrating and destroying the wildlife flora are a global problem. That fact, combined with the open trade of people, materials and plants creates the need to address this problem in a global scale.

Of course one team of 15 people cannot do that by themselves. It is thus proposed that task force centres are established in every continent, with permanent personnel, fully funded by the income of our agrodiagnostics section of the project and with the approval of every respective country.

These centres will be in charge of receiving, distributing and conducting the diagnostics test, while gathering data on each pathogens epidemiology within the country but in neighbouring states as well. Along with governments, they will work together to draft new conservation policies and promote the importance of environmental conservation in their local societies.

After meetings with members of organisations that include kindred taskforces in their organisational plan, we reached the conclusion that each centre would require at least the following:

  • A General Manager
  • A scientific team of at least 10 people in charge of testing
  • A team in charge of logistics and transportation
  • A team of at least 2 data analysts
  • A legal team of at least 3 people
  • A team in charge of communication and public media engagement
  • An obligatory diverse team acting as the liaisons of the taskforce and the general public

Choosing the countries where those taskforces would be set up was not an easy feat as many parameters had to be put under consideration including but not limited to the severity of the state of the local wildlife, the legal prerequisites for realising such a task, the costs of setting up a team there and the ease of access from and to neighbouring countries. For each primary taskforce choice presented in the map below a short explanation is provided:

Elaboration on our primary taskforce locations


Brazil was one of the most abundantly clear candidates for setting up a taskforce due to its extremely rich biodiversity, the importance of its tropical forests worldwide, its location in the continent of South America, its large population and its large contribution in the worldwide plant produce economy.


Italy presents itself as a prime candidate for Mediterranean and Alpine biodiversity with rapid geological switches between ecosystems. Due to its prime location in the centre of Southern Europe, its proximity to Greece and the fact that it is a member of the European Union, Italy is an ideal place for setting up our first outsourced European taskforce.

DR Congo

The Democratic Republic of Congo is one of the most rapidly developing countries of the African continent. It was partly chosen for the same reasons Brazil was, that is, for its prime geographical position and its vast rainforests.


India presents some unique features that little to no countries possess in the globe. Its unique biodiversity, unusual crop produces, extremely rich sociocultural background and the growing public interest on environmental conservation strategies make it a place in the project’s portfolio that can widen possible target plants while being a testing ground for locating the very much needed balance between agrodiagnostics and conservation diagnostics.


Australia is a vast country that entails many unique ecosystems ranging from desserts to rainforests. Due to its unusual fauna it comprises a major source of information on the constant correlations and interactions between flora and fauna and how our efforts on plant conservation may aid animal conservation as well. Moreover, Australian laws are extremely favourable in matters that revolve around the preservation of their ecosystems.

Important side-notes

Minimal Waste

Our team is very keen on leading and assisting conservation efforts around the world. It is thus only natural that we lead by example. Our test kit as well as the delivery methods that we will employ are expected to be the less harmful to the environment. For instance, we opted in using biodegradable plastic materials made from corn rather than classic plastic. Moreover, once our project is scaled up, we aim to use 3D printers powered by sunlight to mass produce our testing platforms. In addition to that, when the aforementioned taskforces are fully set up and running, the kits arriving them won’t arrive separately but in bulk instead. For example, bulk ethanol and nanopure water could be delivered in them and the proper dilutions for the solutions needed could be performed by the personnel there. This saves a lot of transportation costs while also minimising the lab consumables used.

Alternatively, transportation costs and general upkeeping costs could be minimised by establishing close collaborations with local business in matters like supplies or server upkeeping for the storage of the collected data.


Our project during its deployment must be validated and verified with two ways; one in a scientific environment and one in the sociopolitical environment. The first one is an almost de facto prerequisite for the kickstarting of our project and the acquisition of the stakeholders. The later must come through meetings with the powers that be in multiple governing levels. We understand that this is no task for an undergraduate team yet we remained convinced that the seriousness of our efforts along with support from our parent institutions [University of Crete and Foundation for Research and Technology Hellas (F.O.R.T.H)] will be enough of a motoring force for initialising this part of our project.


Any conservation effort, regardless of its origin, is almost doomed to fail if it’s not backed up by corresponding laws from each country. Furthermore we understand that our proposal will not be a panacea in this problem. Additionally its full implementation, if everything goes according to plan, will not happen before 2032. 10 years, in the context of environmental conservation, is a lot of time. By then more, and likely better, solutions will exist that, in addition to ours, will make tackling the problem even easier.

In this effort we are not alone. We aim to help kindred efforts to ours as well by using the potency of our proposal to push for better law and policy making. Our solution is global and so the conservation laws should be. By exhibiting diagnostic consistency, a large spectrum of target pathogens and proper data collecting and analysis, we reckon that we can indeed shift the world towards adopting a universal set of policies.


1. Tsugama, D., Liu, S. & Takano, T. A rapid chemical method for lysing Arabidopsis cells for protein analysis. Plant Methods 7, 22 (2011).

2. Emaus, M.N., Cagliero, C., Gostel, M.R. et al. Simple and efficient isolation of plant genomic DNA using magnetic ionic liquids. Plant Methods 18, 37 (2022).

3. BIO Intelligence Service (2011), Disturbances of EU forests caused by biotic agents, Final Report prepared for European Commission (DG ENV)

4. A.E.S. Cerqueira, T.H. Silva, A.C.S. Nunes, D.D. Nunes, L.C. Lobato, T.G.R. Veloso, S.O. De Paula, M.C.M. Kasuya, C.C. Silva, Amazon basin pasture soils reveal susceptibility to phytopathogens and lower fungal community dissimilarity than forest, Applied Soil Ecology,Volume 131, 2018, Pages 1-11, ISSN 0929-1393,

5. Parrella, G., Gognalons, P., Gebre-Selassiè, K., Vovlas, C., Marchoux, G., 2003. An update of the host range of tomato spotted wilt virus. Journal of Plant Pathology, 85(4 Special issue), 227-264. doi: 10.2307/41998156

6. Turina, M., Tavella, L., Ciuffo, M., 2012. Tospoviruses in the Mediterranean area. Advances in Virus Research, 84, 403-437. doi: 10.1016/B978-0-12-394314-9.00012-9