Difference between revisions of "Team:Oxford/Applied Design"

Line 77: Line 77:
 
     <div style="margin-top: 600px"></div>
 
     <div style="margin-top: 600px"></div>
  
−
<h2 id='title'>Why are synthetic biology diagnostics useful?</h2>
+
<h2 id='title'>Introduction</h2>
−
<p>Conventional diagnostics are currently limited by factors such as resource availability and cost. Synthetic biology provides an opportunity for existing sophisticated biological designs to be exploited and integrated into new systems. Multiplexed signal processing allows for dynamic processing of multiple diagnostic variables, aiding precise health care decisions therefore directly benefiting doctors and patients. Importantly, this form of biotechnology is far more cost-effective and can support developing areas with poorer infrastructure. We therefore believe that synthetic biology diagnostics lie at the heart of the future of medicine.</p><br>
+
<p>iGEM encourages all teams to take their projects beyond the lab and to take a holistic approach to design. The question “What is our real world problem?” has been a key consideration from the beginning, and has guided our project throughout the summer.</p><br>
 +
<p>To put our diagnostic device into context, we considered various aspects including safety, accessibility and socioeconomic factors in Latin America. Many design iterations were built upon over the course of the summer, influenced by discussions with experts from a range of disciplines, including blood coagulation, microfluidics and general diagnostic devices.</p><br>
 +
<p>Having thoroughly examined and evaluated various design options, we propose a final design for our system which fulfils our criteria for a suitable diagnostic device.</p><br>
  
−
<h2>Why did we focus on diagnostics?</h2>
+
<h2>What diagnostics for Chagas disease currently exist?</h2>
  
−
<p>Very early on, we each came up with an idea for our iGEM project and presented it to the group. You can see some of these on our <a href= "https://2017.igem.org/Team:Oxford/InitialIdeas">Initial Ideas page</a>.</p><br>
+
<p>Diagnosis of Chagas disease is difficult, as the disease is mostly asymptomatic in the acute phase and for the majority of the chronic phase.</p><br>
−
<p>We carried out a public survey in the UK, where more than half of the 200 surveyed wanted a synthetic biology solution for disease diagnosis. You can read more about our surveys on our <a href= "https://2017.igem.org/Team:Oxford/HP/Silver">Silver Human Practices page</a>.<br>
+
−
<br><p>We identified a gap in the field of rapid, point-of-care diagnostics which arises when antibody-based technologies cannot be used, for example diagnosis of diseases in infants or immunocompromised patients. As a result, we decided to use the flexibility and versatility of synthetic biology to design a platform technology which addresses these issues.
+
−
</p></br>
+
  
−
<h2>What is Chagas disease?</h2>
+
<p>We spoke to Dr Carol Lole Harris, who advised us on the practical and social considerations involved in the application of our diagnostic in Latin America, based on her experience working in Paraguay. We discussed the current tests available for Chagas and Carol emphasised that a spot test would be by far the best option for a multitude of reasons.</p><br>
−
<p>Our cell-free diagnosis kit is designed to diagnose Chagas disease in its acute phase using a simple blood test. Chagas disease is a neglected tropical disease endemic to Latin America that impacts 6-7 million people, of whom 95% lack sufficient diagnosis or treatment. We decided to focus our efforts on designing a diagnostic for congenital Chagas disease, since current point-of-care diagnostics cannot be used to detect Chagas disease in infants. Current treatments using benznidazole and nifurtimox are almost 100% effective if given shortly after the onset of the acute phase. However, lack of diagnosis leads to the onset of the chronic phase, which causes irreversible pathological consequences to the heart, digestive system, and nervous system. We hope to make a positive contribution towards this cause with our project. </p><br>
+
−
<p> You can read more about this on our <a href= "https://2017.igem.org/Team:Oxford/Chagas_Disease">Chagas disease page</a>.</p><br>
+
  
−
<!-- insert Chagas page logo -->
+
<p>Following further reading, we created a table of our findings to highlight the lack of a rapid and feasible diagnostic for congenital Chagas disease. </p><br>
  
−
<h2>What is our solution?</h2>
 
−
<p>We have designed two systems - one DNA based and one protein-based - to detect a protease, cruzipain. Cruzipain is produced and secreted  by <em>T. cruzi</em>  in the blood and has a specific cleavage sequence, which is ideal for the input. Our systems have bivalirudin as the output for both methods. Bivalirudin is a small peptide that acts as an anticoagulant. Therefore if bivalrirudin were produced in response to the presence of cruzipain, the blood would be inhibited from clotting. These systems are designed to be cell-free and freeze-dried to ensure safety and ease of transport, before being added to a sample of blood.</p><br>
 
  
−
<p>For our DNA-based system, we have designed a TetR molecule with a cleavage site for TEV protease. Our TetR will start bound to its DNA operator, repressing the production of an output protein. When it is cleaved by TEV, repression is relieved, and the reporter produced.</p><br>
+
<h6>Table 1: Main diagnostic methods currently used to diagnose Chagas disease</h6>
 +
<table class="table table-hover">
 +
    <thead>
 +
      <tr>
 +
        <th>Test</th>
 +
        <th>How it works</th>
 +
        <th>Benefits</th>
 +
        <th>Limitations</th>
 +
        <th>Suitable for newborns</th>
 +
      </tr>
 +
    </thead>
 +
    <tbody>
 +
      <tr>
 +
        <td>Whole parasite microscopy</td>
 +
        <td>
 +
        <ul>
 +
        <li>Preparation of Giesma blood smears</li>
 +
        <li>Visualised using light microscopy</li>
 +
        </ul>
 +
        </td>
 +
        <td>
 +
        <ul>
 +
        <li>Established method</li>
 +
        <li>Carried out by already trained professionals</li>
 +
        </ul>
 +
        </td>
 +
        <td>
 +
        <ul>
 +
        <li>Not suitable for the when there is little <em>T. cruzi</em> in the blood</li>
 +
        <li>Not always possible to differentiate between <em>T. cruzi</em> from <em>T. rangeli</em>, which does not cause disease in humans.</li>
 +
        </ul>
 +
        </td>
 +
        <td>Yes</td>
 +
      </tr>
 +
      <tr>
 +
    <td>Polymerase Chain Reaction (PCR)</td>
 +
    <td>
 +
    <ul>
 +
    <li>Molecular detection of <em>T. cruzi</em> DNA is performed using a combination of three real-time PCR assays.</li>
 +
    <li>Acceptable specimen types are EDTA blood, heart biopsy tissue or cerebrospinal fluid.</li>
 +
    </ul>
 +
    </td>
 +
    <td>
 +
    <ul>
 +
    <li>Allows high sensitivity in the acute phase</li>
 +
    <li>Allows the presence of <em>T.cruzi</em> to be accurately distinguished from <em>T. rangeli</em></li>
 +
    <li>Allows direct detection of infection and easy interpretation of results</li>
 +
    </ul>
 +
    </td>
 +
    <td>
 +
    <ul>
 +
    <li>High variation in accuracy and lack of international quality controls</li>
 +
    <li>High cost and complexity means it is not practical to use in a clinical practice</li>
 +
    <li>Further validation is needed to prove whether PCR is suitable to diagnose the  chronic phase of Chagas</li>
 +
    </ul>
 +
    </td>
 +
    <td>Yes</td>
 +
      </tr>
 +
      <tr>
 +
      <td>Serological tests</td>
 +
      <td>
 +
      <ul>
 +
      <li>Detection of antibodies against <em>T. cruzi</em></li>
 +
      <li>Includes techniques such as indirect fluorescent antibody (IFA) test, a commercial enzyme immunoassay (EIA) and immunochromatographic tests</li>
 +
      </ul>
 +
      </td>
 +
      <td>
 +
      <ul>
 +
      <li>Can be used for acute phase and chronic phase</li>
 +
      <li>High specificity and sensitivity</li>
 +
      <li>Commercialised and approved for use by WHO</li>
 +
      <li>Low-cost formats are available</li>
 +
      </ul>
 +
      </td>
 +
      <td>
 +
      <ul>
 +
      <li>Cross reactivity can occur with diseases, such as leishmaniasis and schistosomiasis </li>
 +
      <li>Performance of these tests is lower than reported by their manufacturer, especially against specific strains of <em>T. cruzi</em></li>
 +
      <li>Not suitable for immunocompromised patients and newborns</li>
 +
      </ul>
 +
      </td>
 +
      <td>No</td>
 +
  </tr>
 +
    </tbody>
 +
  </table>
 +
<br>
  
−
<p>For our protein-based system, we have designed an amplificatory protein circuit encased in outer membrane vesicles (OMVs). Both our input (cruzipain) and our intermediate output (TEV protease) are proteases. The amplification components of our system is a split TEV protease, the two halves of which are made accessible to dimerise in the presence of cruzipain. Upon dimerisation, the protease is activated and can go on to activate more of itself in an amplificatory positive feedback loop. Active TEV protease can then cleave and release bivalirudin, which acts as the <a href = "https://2017.igem.org/Team:Oxford/Design#C3">reporter</a> of our system by inhibiting blood clotting.</p><br>
+
<p>We also spoke to Professor Dias Borges Lalwani is professor of epidemiology at the Faculty of pharmaceutical sciences at UFAM, whose contact details were forwarded to us by the AQA Amazonas team. We were fortunate enough to be able to arrange a Skype meeting with Jaila, which helped us contextualise the problem of Chagas highlighting the difficulty of seeking diagnosis given non-specific early disease symptoms. </p><br>
  
−
<p> You can read more about this on our <a href= "https://2017.igem.org/Team:Oxford/Design">Design page</a>.</p>
+
<h2>Why congenital Chagas disease?</h2>
 +
<p>We spoke to Dr Alonso-Vega of the University of San Simón, Cochabamba who ran the National Congenital Chagas Disease Programme in Bolivia from 2004-2009, and is an expert in congenital Chagas. Dr Alonso-Vega’s Chagas Disease Program recommended the need for a new congenital diagnostic. She gave us specific guidance for implementation of our diagnostic device in Bolivia and confirmed the suitability of our congenital test in Bolivian hospitals, where most deliveries occur.</p><br>
  
−
<!-- insert project design page logo & link to right of text -->
+
<p>We first contacted Professor Yves Carlier early on in our project to gain more information about the pathology of congenital Chagas disease. Professor Carlier is a researcher in infectious diseases and clinical immunology at the Université Libre de Bruxelles. He re-emphasised the need for a diagnostic for neonates, strengthening our resolve to focus on congenital Chagas disease. He also informed us about the benefits of diagnosing Chagas disease in neonates, such as that treatment with benznidazole cures around 100% of babies if given before one year of age but not later in life.</p><br>
  
−
<h2>What is our strategy?</h2>
+
<p>We also learnt that Chagas treatment for under 15’s is free, giving us confidence that our diagnostic would be accessible and impactful in containing Chagas if provided in a hospital setting. Our discussion with her further emphasised the requirement for a rapid point of care test that would allow treatment of infected newborns to begin before they left the hospital.</p><br>
  
−
<h3>Wet Lab</h3>
+
<h2><a href = "https://2017.igem.org/Team:Oxford/Applied_Design_Developing">How did we develop our design?</a></h2>
−
<p>For our DNA-based system, we characterised the pTet + eYFP part using fluorescence microscopy and plate reading, which showed that TetR can bind to the pTet and repress the output fluorescence significantly. This part has a carefully picked ribosome binding site and promoter strength to optimise our system for minimal false positives and negatives when eYFP is replaced with TEV protease production. Hence it was highly important to detect how repression was relieved when Anydrotetracyline(ATC) was introduced, which acts on TetR.</p><br>
+
<p>Based on our findings from the OpenPlant conference, we established a set of criteria for our applied design - the 4 E’s (‘Effectiveness’, ‘Ease of use’, ‘Economics’ and ‘Environment & Safety’).</p><br>
  
−
<p>The sfGFP+Quencher was characterised for our OMV system. This part was critical to identify if sfGFP (GFP modified to fold in the periplasm) can be quenched by a quenching peptide linked with a protease specific cleavage sequence. We tested the functionality and sensitivity of the part to TEV protease through a double transformation of the part and TEV plasmids. Plate reader and fluorescence microscopy on this part identified that the Quencher can quench sfGFP fluorescence and that quenching can be relieved by introducing the TEV protease.</p><br>
+
<p>Prototyping involved extensive consideration of various design iterations. We progressed from paper to cardboard to 3D printing - at each stage of this process, we were able to iron out flaws in our design and to incorporate new features from our evaluations.
  
−
<p> You can read more about this in our <a href= "https://2017.igem.org/Team:Oxford/Overview_Wet_Lab">Wet Lab section</a>.</p>
 
  
 +
<p>Our current kit meets our criteria established from our 4E’s framework: it is effective, easy-to-use, economically viable and environmentally safe. Furthermore, it incorporates many of the recommendations provided to us by the experts we contacted. Our solution is a rapid, point-of-care diagnostic for congenital Chagas disease which can easily be used by any healthcare professional.</p><br>
  
−
<h3>Real-world perspectives</h3>
+
<p>Following extensive design and development, we produced a 3D prototype version of our kit. We spoke to Tim Ring, the vice president of safe-tec sales and marketing who generously sent us MICROSAFE® pipette samples so we could test the compatibility of these with our kit design. These pipettes are advantageous for blood collection and allowed us to test our applied design more rigorously.</p><br>
−
<p>Our project has been guided throughout by input from experts in Latin America and medical professionals in the UK. Conversations with the public during our outreach activities also helped us to consider perspectives around synthetic biology outside the lab. You can read more about this on our <a href= "https://2017.igem.org/Team:Oxford/HP/Gold_Integrated">Gold & Integrated Human Practices page</a> and <a href= "https://2017.igem.org/Team:Oxford/Engagement">Education & Public Engagement page</a>.</p><br>
+
  
−
<p>Consultation with relevant stakeholders, including HeLEX (Centre for Health, Law and Emerging Technologies), InSIS (Institute for Science Innovation and Society) and numerous experts worldwide, has helped to inform ethical and social considerations relevant to our project. These consultations have directly fed back into our applied design to enable a bedside-to-bench approach helping us to design and prototype a diagnostic kit for Chagas disease which is easy-to-use, cheap to manufacture and has minimal risk to the environment. <a href= "https://2017.igem.org/Team:Oxford/Applied_Design">You can read more about this on our Applied Design page</a>.</p><br>
+
<p> Our 3D prototype design was refined by the healthcare professionals we contacted for evalutation. The insights from Sarah Dragonetti (Registered Nurse) and Dr. Ben Riley (General Practice doctor) were hugely useful in assessing and refining the practicality of our diagnostic kit, consequently improving our applied design.</p>
  
−
<p>To support the integration of our device into existing healthcare systems, our dialogue with HeLEX inspired us to create a policy proposal to address gaps in regulation present in current infrastructure.</p><br>
 
  
−
<a href="https://2017.igem.org/Team:Oxford/Chagas_Public_Policy"><img class="img-responsive img-center" width="200px;" src="https://static.igem.org/mediawiki/2017/9/98/T--oxford--chagas_disease--button.png"></a><br>
+
<h2><a href = "https://2017.igem.org/Team:Oxford/Applied_Design_Implementation">How will we implement our design?</a></h2>
  
−
<p>Our cell-free design has been inspired by consultation with Dr Keith Pardee. Combining this with our discussions about safety with Piers Millet and HeLEX, we designed our parts for the wet lab with this in mind and produced a report outlining the barriers faced by cell-free technology. We hope this will prove useful for future iGEM teams using cell-free technology.</p><br>
+
<p>We consider integration of our device into existing healthcare systems and current infrastructure a key challenge, and therefore a fundamental aspect of our applied design considerations.</p><br>
  
−
<a href="https://2017.igem.org/Team:Oxford/Cell_Free_Report"><img class="img-responsive img-center" width="200px;" src="https://static.igem.org/mediawiki/2017/c/ca/T--oxford--cellfreereport.png"></a>
+
<p>We received invaluable advice from David Sprent, an expert in International Supply Chain, and Juan Solano and Alfons Van Woerkom, representatives of the Global Fund who advised us on evaluating costs of manufacture, transport and taxation of a kit for end use by the Bolivian healthcare system. </p><br>  
−
<br>
+
  
−
<h3>Modelling</h3>
+
<p>We consulted with HeLEX, (Centre for Health, Law and Emerging Technologies) and with InSIS (Institute for Science, Innovation and Society). InSIS researches and informs key contemporary and emerging issues and processes of social, scientific, and technological change. The insights from both HeLEX and InSIS were highly useful in evaluating the ethical and social issues related to our project, and heavily influenced our applied design.</p></br>
−
<p> Modelling was an inseparable part of our design process: it allowed us to quickly test our theoretical designs and identify key design parameters that could improve our design. We worked closely with experts throughout developing our models. Collaborations have allowed us to refine our methodology by applying it to the different systems of other teams, inspiring us to document it to help future teams. <a href= "https://2017.igem.org/Team:Oxford/Model">You can read more about this in our Modelling section</a>.</p><br>
+
  
−
<p>We were able to model the impact our diagnostic would have on the epidemiology of Chagas disease in Bolivia by working closely with Professor Michael Bonsall (a mathematical biologist) and Dr Yves Carlier (a Chagas epidemiologist) to create a disease model that we hope to publish later this year. <a href= "https://2017.igem.org/Team:Oxford/Disease_Model">You can read more about this on our Disease Modelling page</a>.</p><br>
+
<p>Professor Keith Pardee has worked on cell-free technologies, and he informed us how cell-free systems could solve a lot of issues around safety and cost.</p><br>
  
−
<h2>What are our visions for the future?</h2>
+
<p>Dr Piers Millett is a Senior Research Fellow at the Future of Humanity Institute, and he introduced us to the concept of platform technologies and how this could be applied to our project. This provides a good way of fast tracking future developments because the platform will already have met regulatory approval, therefore allowing the device to be more rapidly adapted for other diseases.></p><br>
−
<h3>Experiments we want to carry out</h3>
+
−
<p>To develop our system into something which can undergo clinical trials and hopefully become a successful product, we have a vision for the experiments that need to be performed. These are detailed at the end of our Results pages - <a href= "https://2017.igem.org/Team:Oxford/Results_DNA">DNA-based</a> and <a href= "https://2017.igem.org/Team:Oxford/Results_Protein">Protein-based</a>.</p><br>
+
−
 
+
−
<p>For our DNA-based system, we envision the progression from a proof-of-concept system to gradually introducing each ‘real’ components, and testing that this does not perturb our system and corroborates our modelling. Additionally, we wish to check the efficacy of different lysates and the freeze-drying process.</p><br>
+
−
 
+
−
<p>For our protein-based system, the aim is to first express our components in outer membrane vesicles, before trialing methods of lysing the OMVs and assaying the efficacy of the split-TEV protease molecule.</p><br>
+
−
 
+
−
<h3>Future visions for our kit</h3>
+
−
<p>We have designed a software tool to facilitate further applications of our project, as our system may be applied to a range of diseases. This is an open-source tool so that researchers may add to a growing database of pathogens and specific protease cleavage sites.</p><br>
+
−
 
+
−
<p>As our kit is modular, it can be easily and cheaply adapted to diagnose different diseases: the cost of changing the disease is then only the input block, not also the output block. Our vision for the future is that a streamlined manufacturing process can be established for rapid development of new diagnostic modules as more specific proteases are characterised and validated as biomarkers.</p><br>
+
−
 
+
−
<a href= "https://2017.igem.org/Team:Oxford/Software">You can see our Software Tool here</a>.<br>
+
  
 
<h2>References</h2>
 
<h2>References</h2>
  
−
<p>Courbet A., Renard E., and Molina F. 2016 Bringing next‐generation diagnostics to the clinic through synthetic biology. EMBO Mol Med 8: 987–991</p>
+
<p>Carlier, Y. and Truyens, C. 2017 Maternal-fetal transmission of Trypanosoma cruzi. Second Edition, American Trypanosomiasis Chagas Disease: One Hundred Years of Research: Second Edition. Second Edition. Elsevier Inc. doi: 10.1016/B978-0-12-801029-7.00024-1.</p>
−
 
+
−
<p>Slomovic S., Pardee K., and Collins J.J. 2015 Synthetic biology devices for in vitro and in vivo diagnostics. Proc Natl Acad Sci USA 112: 14429–14435.</p>
+
−
 
+
−
<p>Wehr, M. C. et al. 2006 ‘Monitoring regulated protein-protein interactions using split TEV’, Nat Meth, 3(12), pp. 985–993. Available at: http://dx.doi.org/10.1038/nmeth967.</p>
+
  
−
<p>Alves, N. J. et al. 2016 ‘Protecting enzymatic function through directed packaging into bacterial outer membrane vesicles’, Scientific Reports. Nature Publishing Group, 6(1), p. 24866. doi: 10.1038/srep24866.</p>
+
<p>Cencig, S. et al. 2012 ‘Evaluation of benznidazole treatment combined with nifurtimox, posaconazole or AmBisome?? in mice infected with Trypanosoma cruzi strains’, International Journal of Antimicrobial Agents. Elsevier B.V., 40(6), pp. 527–532. doi: 10.1016/j.ijantimicag.2012.08.002.</p>
  
 
</div>
 
</div>

Revision as of 02:44, 2 November 2017


APPLIED DESIGN

Introduction

iGEM encourages all teams to take their projects beyond the lab and to take a holistic approach to design. The question “What is our real world problem?” has been a key consideration from the beginning, and has guided our project throughout the summer.


To put our diagnostic device into context, we considered various aspects including safety, accessibility and socioeconomic factors in Latin America. Many design iterations were built upon over the course of the summer, influenced by discussions with experts from a range of disciplines, including blood coagulation, microfluidics and general diagnostic devices.


Having thoroughly examined and evaluated various design options, we propose a final design for our system which fulfils our criteria for a suitable diagnostic device.


What diagnostics for Chagas disease currently exist?

Diagnosis of Chagas disease is difficult, as the disease is mostly asymptomatic in the acute phase and for the majority of the chronic phase.


We spoke to Dr Carol Lole Harris, who advised us on the practical and social considerations involved in the application of our diagnostic in Latin America, based on her experience working in Paraguay. We discussed the current tests available for Chagas and Carol emphasised that a spot test would be by far the best option for a multitude of reasons.


Following further reading, we created a table of our findings to highlight the lack of a rapid and feasible diagnostic for congenital Chagas disease.


Table 1: Main diagnostic methods currently used to diagnose Chagas disease
Test How it works Benefits Limitations Suitable for newborns
Whole parasite microscopy
  • Preparation of Giesma blood smears
  • Visualised using light microscopy
  • Established method
  • Carried out by already trained professionals
  • Not suitable for the when there is little T. cruzi in the blood
  • Not always possible to differentiate between T. cruzi from T. rangeli, which does not cause disease in humans.
 Yes
Polymerase Chain Reaction (PCR)
  • Molecular detection of T. cruzi DNA is performed using a combination of three real-time PCR assays.
  • Acceptable specimen types are EDTA blood, heart biopsy tissue or cerebrospinal fluid.
  • Allows high sensitivity in the acute phase
  • Allows the presence of T.cruzi to be accurately distinguished from T. rangeli
  • Allows direct detection of infection and easy interpretation of results
  • High variation in accuracy and lack of international quality controls
  • High cost and complexity means it is not practical to use in a clinical practice
  • Further validation is needed to prove whether PCR is suitable to diagnose the chronic phase of Chagas
 Yes
Serological tests
  • Detection of antibodies against T. cruzi
  • Includes techniques such as indirect fluorescent antibody (IFA) test, a commercial enzyme immunoassay (EIA) and immunochromatographic tests
  • Can be used for acute phase and chronic phase
  • High specificity and sensitivity
  • Commercialised and approved for use by WHO
  • Low-cost formats are available
  • Cross reactivity can occur with diseases, such as leishmaniasis and schistosomiasis
  • Performance of these tests is lower than reported by their manufacturer, especially against specific strains of T. cruzi
  • Not suitable for immunocompromised patients and newborns
 No

We also spoke to Professor Dias Borges Lalwani is professor of epidemiology at the Faculty of pharmaceutical sciences at UFAM, whose contact details were forwarded to us by the AQA Amazonas team. We were fortunate enough to be able to arrange a Skype meeting with Jaila, which helped us contextualise the problem of Chagas highlighting the difficulty of seeking diagnosis given non-specific early disease symptoms.


Why congenital Chagas disease?

We spoke to Dr Alonso-Vega of the University of San Simón, Cochabamba who ran the National Congenital Chagas Disease Programme in Bolivia from 2004-2009, and is an expert in congenital Chagas. Dr Alonso-Vega’s Chagas Disease Program recommended the need for a new congenital diagnostic. She gave us specific guidance for implementation of our diagnostic device in Bolivia and confirmed the suitability of our congenital test in Bolivian hospitals, where most deliveries occur.


We first contacted Professor Yves Carlier early on in our project to gain more information about the pathology of congenital Chagas disease. Professor Carlier is a researcher in infectious diseases and clinical immunology at the Université Libre de Bruxelles. He re-emphasised the need for a diagnostic for neonates, strengthening our resolve to focus on congenital Chagas disease. He also informed us about the benefits of diagnosing Chagas disease in neonates, such as that treatment with benznidazole cures around 100% of babies if given before one year of age but not later in life.


We also learnt that Chagas treatment for under 15’s is free, giving us confidence that our diagnostic would be accessible and impactful in containing Chagas if provided in a hospital setting. Our discussion with her further emphasised the requirement for a rapid point of care test that would allow treatment of infected newborns to begin before they left the hospital.


How did we develop our design?

Based on our findings from the OpenPlant conference, we established a set of criteria for our applied design - the 4 E’s (‘Effectiveness’, ‘Ease of use’, ‘Economics’ and ‘Environment & Safety’).


Prototyping involved extensive consideration of various design iterations. We progressed from paper to cardboard to 3D printing - at each stage of this process, we were able to iron out flaws in our design and to incorporate new features from our evaluations.

Our current kit meets our criteria established from our 4E’s framework: it is effective, easy-to-use, economically viable and environmentally safe. Furthermore, it incorporates many of the recommendations provided to us by the experts we contacted. Our solution is a rapid, point-of-care diagnostic for congenital Chagas disease which can easily be used by any healthcare professional.


Following extensive design and development, we produced a 3D prototype version of our kit. We spoke to Tim Ring, the vice president of safe-tec sales and marketing who generously sent us MICROSAFE® pipette samples so we could test the compatibility of these with our kit design. These pipettes are advantageous for blood collection and allowed us to test our applied design more rigorously.


Our 3D prototype design was refined by the healthcare professionals we contacted for evalutation. The insights from Sarah Dragonetti (Registered Nurse) and Dr. Ben Riley (General Practice doctor) were hugely useful in assessing and refining the practicality of our diagnostic kit, consequently improving our applied design.

How will we implement our design?

We consider integration of our device into existing healthcare systems and current infrastructure a key challenge, and therefore a fundamental aspect of our applied design considerations.


We received invaluable advice from David Sprent, an expert in International Supply Chain, and Juan Solano and Alfons Van Woerkom, representatives of the Global Fund who advised us on evaluating costs of manufacture, transport and taxation of a kit for end use by the Bolivian healthcare system.


We consulted with HeLEX, (Centre for Health, Law and Emerging Technologies) and with InSIS (Institute for Science, Innovation and Society). InSIS researches and informs key contemporary and emerging issues and processes of social, scientific, and technological change. The insights from both HeLEX and InSIS were highly useful in evaluating the ethical and social issues related to our project, and heavily influenced our applied design.


Professor Keith Pardee has worked on cell-free technologies, and he informed us how cell-free systems could solve a lot of issues around safety and cost.


Dr Piers Millett is a Senior Research Fellow at the Future of Humanity Institute, and he introduced us to the concept of platform technologies and how this could be applied to our project. This provides a good way of fast tracking future developments because the platform will already have met regulatory approval, therefore allowing the device to be more rapidly adapted for other diseases.>


References

Carlier, Y. and Truyens, C. 2017 Maternal-fetal transmission of Trypanosoma cruzi. Second Edition, American Trypanosomiasis Chagas Disease: One Hundred Years of Research: Second Edition. Second Edition. Elsevier Inc. doi: 10.1016/B978-0-12-801029-7.00024-1.

Cencig, S. et al. 2012 ‘Evaluation of benznidazole treatment combined with nifurtimox, posaconazole or AmBisome?? in mice infected with Trypanosoma cruzi strains’, International Journal of Antimicrobial Agents. Elsevier B.V., 40(6), pp. 527–532. doi: 10.1016/j.ijantimicag.2012.08.002.