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. 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.
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.
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.