Difference between revisions of "Team:Munich"

Revision as of 10:16, 30 August 2017

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. 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. Furthermore, Cas13a allows the fast design of target sequences, making our system adaptive to the emergence of new viral outbreaks or fast mutating pathogens.

<b>Step 1:</b><br>
A sample, such as blood or saliva, is placed into the device.

Step 1:
A sample, such as blood or saliva, is placed into the device.

Step 3:
Cas13a binds the target sequence and cuts it in smaller fragments. After this inital digestion, the enzyme changes into a RNAase-active conformation.

Step 5:
An easy to interpret read-out tells whether a specific pathogen was present in the sample.

<b>Step 2:</b><br>
RNA is purified with the help of our reusable, 3D-printed microfluidic device.

Step 2:
RNA is purified with the help of our reusable, 3D-printed microfluidic device.

Step 4:
While in this conformation, Cas13a degrades the read-out RNA, producing a visible signal.

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-      CascAID stands for Cas13a Controlled Assay for Infectious Diseases. Our goal this year is to create a novel paper-based microfluidic device for the detection of specific RNA sequences.
+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. 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. Furthermore, Cas13a allows the fast design of target sequences, making our system adaptive to the emergence of new viral outbreaks or fast mutating pathogens.
-      Nowadays, there is a trend among medical practitioners to prescribe antibiotics when bacterial infections are suspected without a laboratory confirmation as a way for speeding up recovery. This has led to an increase in antibiotic-resistant bacteria that makes it difficult to treat infected patients. Nucleic acid-based detection methods could allow for faster diagnosis. Therefore, we seek to develop a method that allows to report the presence of a specific pathogen within hours. For this reason, we will use the CRISPR effector Cas13a, which is able to target specific single-stranded RNA. This system allows for the simple and fast design of new sequence targets, being an ideal tool for detecting fast mutating pathogens. In the long term, we see our device as an easy-to-use and fast diagnostic tool in developing countries as well as an instrument in developed countries for discerning between bacterial and viral infections, that could help reducing antibiotics prescription.
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