RIBOTOX

Engineering

Implementation

Our kit is a multi-use rapid preeclampsia kit. With millions of people suffering from preeclampsia every year, it has come to our attention that a cheap and quick detection kit had to be made. Pre-existing detection methods are either extremely inaccurate (eg: pressure cuffs) or quite expensive and slow. Therefore, our team focused on increasing the accuracy, reducing the cost, and shortening the time required to take the test.

The aspects of our kit:

Biology

The synthetic biology component of our kit is the most vital, and we created three generations of toehold switches to this end. To read more about our toehold switches design, go to our design page, to learn more about testing them, go to our measurements and results page.

The toehold switch which we would use in our kit is our best, and final iteration - gen3. This toehold switch is an AND-gate switch which can detect multiple miRNA triggers by binding them together with synthetic anti-miRNAs to form larger trigger complexes. We tested this switch in the lab to show it can discriminate between a ratio of concentrations typical between patients with and without Preeclampsia. If our kit was implemented, we would conduct a large research study to find even more miRNAs upregulated in patients with Preeclampsia, and then create more than 2 input AND gates based on the AND gate framework we showed works with this switch.

In order to ensure patients’ serum has a concentration of miRNA high enough for us to detect the percentage difference, we have designed an isothermal amplification strategy for our miRNA triggers, involving miRPA and RPA. Read more about these on our Design page.

Hardware

On top of our wet lab, we extensively researched methods to measure luciferase output from our designed circuits.

We designed a magnetic bead-based miRNA extraction system, which could be further developed to be used in clinical trials to improve specificity and accuracy of our extraction system. In addition, we developed and assembled a luminometer circuit, which measures luminescence to return a positive or negative result. If our hardware was to be implemented, this luminometer design can be adapted to rollout a low-cost, high-specificity luminometer.

Stages of kit implementation

For a extensive rollout of our proposed detection kits, the following stages of work would have to be completed:

Fundraising and project proposal
- For mass production and professional research, sums in the millions of pounds would be required. In order to achieve this fundraising requirement, attracting investment through project proposals is key.
Research
- We can use the AND gate framework to create multi-input toehold switches which detect a much larger array of miRNAs for higher specificity.
- More research in order to reduce false results, minimise errors, and maximise specificity is required.
Designing toehold switches and probes for miRPA using our software tools and the general project framework
- Once preliminary research is completed, the first phase of designing the kit begins.
- Once AND gate toehold switch and miRNA probes are designed, the kit is ready to be tested.
Trials, both clinical and non-clinical
- To ensure maximum safety and accuracy of our product, multiple trial phases are required.
- In vitro testing, human testing would be required. One advantage of our product is that our kit has no side effects bar collecting a small sample of blood from the tested individual. This will, therefore, reduce time and cost for clinical trials compared to many other medical equipment.
Rollout
- After completing testing, our kit has to be sold and distributed to pregnancy clinics, hospitals, national healthcare services, and governmental bodies.
- This is further discussed below, where we look at ‘Customer Discovery’

SWOT Analysis:

To carefully judge the pros and cons of our kit, we created a SWOT analysis table.

Strengths

Requires little equipment bar our kit, and highly cost effective, even when rolled out in large numbers (See Cost Model under Modelling)
Diagnosis in under one hour
Pre-Eclampsia product new to market
Serum sample is easy to obtain
Small current market for blood-test- based Preeclampsia detection
Innovative use of AND-gate toehold switches to prevent ‘leakiness’ and false positives and increase accuracy
Preeclampsia test market estimated to be $1.1 billion by 2027 ^[1]

Weaknesses

Absence of creation of large-scale production
Solutions for the test require containment in dry ice
Hardware requires an energy source from either batteries or an outlet plug, making it hard to implement in under-developed areas
Lack of copyright for our test

Opportunities

Currently no other blood test for Pre-Eclampsia
Half the price of screening during first and second trimester
A third of the price of combined early-onset and first trimester screening

Threats

VCGS offers a screening process for first trimester, second trimester, and early-onset preeclampsia
Negative perceptions of genetic engineering from the public

Customer discovery:

Focusing on the different levels of customers that will purchase our kit, we looked at a total of 4 different stages.

From the distributor (us or a manufacturing company):

Customer Level 1:

Governments and NHS: The National Health Service is the publicly funded healthcare system in England, and received a budget of £134 billion in 2019. Furthermore, the NHS delivered 591,759 babies in 2019. Our product would be used by all the expecting mothers who are treated by the NHS to ensure early and reliable diagnosis of pre-eclampsia.

National healthcare systems would purchase the preeclampsia tests for use on expecting mothers in their care.

Customer Level 1.5:

Pharmaceutical TNCs (eg Bayer, GSK)/Chain private-clinics:

National healthcare systems would purchase the preeclampsia tests for use on expecting mothers in their care.

Customer Level 2:

Pharmacies/ Supermarkets:

Pharmacies and Supermarkets would purchase the tests to resell them to their customers that are pregnant and want to test for preeclampsia.

Customer level 3: The Ultimate benefitter

The ultimate benefitter would be the mother who is being tested and her unborn baby as this test would indicate whether treatment is needed and maybe whether an early delivery is needed.

Similar products:

To find competitors in our market, we first looked at similar disease detection methods. Then we found one main competitor, a company who focuses on preeclampsia detection specifically. This competitor, VCGS, provides testing for first and second trimester screening along with early-onset preeclampsia tests. For the first and second trimester tests, the price ranges from $79 to $101. In this case our cost is below the lowest bound which provides a strong incentive to buy from us instead of from VCGS. The early-onset tests (combined with first trimester testing) range in price from $111 to $120. Our product does not directly compete with the early-onset testing services, however, that we are once again below the lower bound of cost is a good sign of our relative price competitiveness. All other competition is not directly against us as all other competitors’ products revolve around placental growth - not a diagnostic blood test like ours. This gives us a smaller and more penetrable market.

Other similar products include Rapid Antigen Tests for travel, PCR tests for COVID-19, and Blood Type testing kits. The rapid antigen tests cost £29 and thus are slightly more cost efficient than our product. However, this price difference is only slight and all of our test is single-use (unlike the RATs), furthermore they test for an entirely different disease. More accurate testing for COVID-19, and another competitor, is by using PCR. These tests cost £79 which makes our test a considerably cheaper technology. Whilst we do not directly compete, since they test for different diseases, this is further evidence for our price being a compelling one. The Blood Type Test Kit is much cheaper than our kit at £7.25. This cost difference can be largely attributed to the difference in hardware pricing. This suggests that our product is not at the cheapest price that could be achievable. However, cost often reduces greatly when mass production is used - due to economies of scale. As such, the price delta is not an issue.

Will governments bulk purchase our kits?

Our kits will replace alternative screening products which currently cost over twice the amount of ours. This provides a cheaper alternative for national health services which are already at their limit of funding, thus providing a strong incentive for governments to bulk purchase our kits. A potential issue could be the requirement for dry-ice shipping and storage. However, through the COVID vaccine rollout, we have seen that healthcare providers have the necessary equipment and capacity to keep a bulk set of tests (namely the Pfizer-BioNTech, and the Moderna vaccines) at less than -15 degrees celsius. The EU, for example, bulk purchased over 900 million Pfizer-BioNTech vaccines^[2] - each which has to be stored at -70 degrees Celsius. Therefore, it is clear that the storage requirements would not negatively impact the chance of governments bulk purchasing our kits. The EU also bought 500 million COVID-19 vaccine does at a cost of £1.75 billion. Conservatively estimating the cost of a single test to be £30, the bloc could purchase 58 million of our tests for the same price. This single level of investment, extrapolating from the number of births in the EU in 2019 (4.2 million)^[3], would provide enough tests for screening of pregnant women for 13.8 years. Preeclampsia is also associated with 30% of overall obstetrics costs^[4] and thus, by buying our product and implementing screening, the associated healthcare providers could save €6.5-9.1 million per country per annum^[5].This shows that the commission, who have a desire to both save lives and reduce costs, would certainly bulk purchase our tests.

Eg: The EU purchased 500 million vaccine doses (Astro Zeneca)with the price being between £3 and £4, total price mounted to about 1.75 billion pounds- 2021^[6].

Key partners and advertising if our kit was to be implemented

To maximise efficiency of our kit production, we must partner up with other companies to purchase supplies and equipment.

Toeholds and probe production will require advice and purchases from gene-specialist companies such as IDT.

Other medical-equipment companies, for example needles to take blood samples, may be supplied from companies like BD.

To promote the purchase of the product, we will have to focus on advertisements. When advertising medical products, it is extremely important to scrutinise its effect on the people who come in contact with the adverts. They must not induce fear or provide false/exaggerated information.

If implemented, the promotion team must research further into laws and regulations around medical equipment and detection kits.

Safety

Despite the invasive nature of our kit (blood sampling), when the blood taking is executed without failure of following protocols (eg: sterile needle, disinfect skin before sampling), there should be no direct side effect.

When looking at the larger picture, our kits may affect the safety of preeclampsia patients when a false negative is shown. This may be due to a production error, anomalous miRNA regulation in the patient, sampling error, or by pure chance. This is in fact the case with most testing kits, especially rapid, low-cost tests, namedly the COVID-19 lateral flow tests. To mitigate the possibility of infringing the safety of preeclampsia patients, we propose the following measures to be taken:

Clearly state to all customers about the chance of false negatives. An example could be:
‘Though our kits provide a rapid testing tool to screen preeclampsia, it is not deterministic. A positive result indicates a high possibility of the testee having preeclampsia. A negative result indicates a high possibility that the testee does not have preeclampsia. When a positive result is obtained, consult a doctor or seek immediate medical aid. When a negative result is obtained, pay attention to potential preeclampsia symptoms, and test again with a doctor at the next obstetrician appointment. This is suggested as false negative results may occur with low probabilities.’
To reduce faulty tests due to contamination, users should be instructed not to physically contact some areas of the testing kit.

Refrences

Cordis.europa.eu. 2021. CORDIS | European Commission. [online] Available at: https://cordis.europa.eu/article/id/429449-safer-pregnancies-thanks-to-quick-easy-preeclampsia-test [Accessed 21 October 2021]
European Commission - European Commission. 2021. Press corner. [online] Available at: https: //ec.europa.eu/commission/presscorner/detail/en/ip_21_2548 [Accessed 21 October 2021].
Ec.europa.eu. 2021. Fertility statistics - Statistics Explained. [online] Available at: https: //ec.europa.eu/eurostat/statistics-explained/index.php?title=Fertility_statistics [Accessed 21 October 2021].
Cordis.europa.eu. 2021. CORDIS | European Commission. [online] Available at: https: //cordis.europa.eu/article/id/429449-safer-pregnancies-thanks-to-quick-easy-preeclampsia-test [Accessed 21 October 2021].
Fox, A., McHugh, S., Browne, J., Kenny, L., Fitzgerald, A., Khashan, A., Dempsey, E., Fahy, C., O’Neill, C. and Kearney, P., 2017. Estimating the Cost of Preeclampsia in the Healthcare System. Hypertension, 70(6), pp.1243-1249.
[Online] Available at: https: //www.cnbc.com/2020/11/17/covid-vaccines-how-much-they-cost-whos-bought-them-and-how-theyre-stored.html [Accessed 21 October 2021].

Team:City of London UK/Implementation