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| − | <font size=7 color=#444444><b>Abstract</b></font>
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| − | The ongoing crisis of increasing antibiotic resistance demands innovative preventive strategies. Recently, the RNA-targeting protein CRISPR-Cas13a has been used for highly sensitive DNA and RNA detection, promising diverse applications in point-of-care diagnostics. We integrated Cas13a in the detection unit of CascAID, our GMO-free diagnostic platform. CascAID combines an automated microfluidic device for rapid lysis and extraction of nucleic acids with a paper-based readout system. We demonstrated the performance of our device by targeting the 16S RNA from E. coli. We improved the detection limit of our platform, using simulations to optimize our amplification scheme and the final readout.
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| − | Conceived as a distributable platform for rapid point-of-care diagnostics, CascAID can be used to distinguish between bacterial and viral infections, thus minimizing the widespread use of antibiotics.
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| − | Furthermore, Cas13a allows the fast design of target sequences, making our system adaptive to the emergence of new viral outbreaks or fast mutating pathogens.
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| | + | <td colspan=6> |
| | + | <div class="captionPicture"> |
| | + | <p> |
| | + | Complete overview of all modular hardware parts in our pathogen detection system. Shown are counterclockwise and starting in in the upper left corner: The Quake valve that controls fluid flow, our sample processing device, the paper strip where a reaction mix is stored and the readout reaction takes place and finally our low-cost fluorescence detector "Lightbringer" that performs the readout measurement. Images are clickable and linked to the corresponding wiki subsection. |
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| − | | + | <font size=7 color=#51a7f9><b style="color: #51a7f9">Hardware</b></font> |
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| | </td> | | </td> |
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| | + | <td colspan = 6 align=center valign=center> |
| | + | <p class="introduction"> |
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| | + | The liberation of diagnostic tests from expensive lab infrastructure requires innovative ways of sample processing and measuring. We therefore developed a set of portable hardware tools with the goal of providing an automated sample-to-answer solution. The heart of our system is <a class="myLink" href="https://2017.igem.org/Team:Munich/Hardware/Detector">‘Lightbringer’</a>, our fluorescence detector, which is capable of measuring kinetics of biological or chemical reactions on <a class="myLink" href= "https://2017.igem.org/Team:Munich/Hardware/Paperstrip" >paper.</a> Built from 3D–printed parts and standard electronic components, it can be assembled for less than 15$, while offering a sensitivity competitive to commercial fluorescence readers. Additionally, tackling the challenge of sample pre-processing in field, we developed a portable <a class="myLink" href="https://2017.igem.org/Team:Munich/Hardware/SampleProcessing"> fluidic system</a>, featuring a temperature control unit for lysis and isothermal PCR. Conceiving a platform independent of lab infrastructure, we demonstrate the feasibility of <a class="myLink" href="https://2017.igem.org/Team:Munich/Hardware/QuakeValve"> controlling fluid flow</a> with bike tires and air balloons. All hardware components are designed and documented with the aim of enabling the community to reproduce and extend our set of tools. |
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