Team:Calgary/Project/BsDetector/DevicePrototype
From 2014.igem.org
Device Prototype
Our team decided that in order to ensure that this system is physically feasible that we would aim to produce a working physical prototype. In this process it was imperative that the end-user was considered and that the materials used made our system economically feasible and compatible with the biological system. Through collaboration with the transformers, detectors and our human practices subteams we were able to manufacture a working prototype as the first stage of development.
Process
(Edit)***Integrate creation paragraph below into this section*** Our device is portable and user friendly. A drop of blood from a patient first needs to go through sample preparation to extract target DNA out of the sample. A sample with extracted DNA then flows into the Polymerase Chain Reaction (PCR) chamber where target DNA is amplified via isothermal PCR. Reaction takes a couple of hours to complete and the sample is manually pushed out of the PCR chamber into separate chambers with B. subtilis. In each of the chambers, B. subtilis is engineered to detect a specific disease allowing for multidiagnosis.
Economic feasability - Two-Tiered Model
Economic feasibility is of the utmost importance when dealing with diagnostics in the developing world and therefore our team chose to tackle this issue using a two-tiered development phase. An inexpensive model would utilize the basics of the colourmetric system in which a simple colour change would indicate presence of the specified disease. This signal is easily visible and readable and requires very little technology other than a chamber for Polymerase Chain Reaction (PCR) to occurs and consequential separation of the sample into separate detection wells. Additionally in order to ensure that their is development potential should this modular system expand to other industries a completely digitized device was also explored. The colourmetric signal of the reporter is picked up by a colour sensor that detects the reflected light of this signal, converts it into a voltage and is output to an LCD screen. This exercises the potential of being able to send results of a test directly to a patient's file with bluetooth use.
Manufacturing
The goal of producing a physical prototype brought different perspectives of the best way to produce such a device. Through expert consultation it was determined that microfluidics would increase the price greatly and would not be ideal for our device, however using polytetrafluoroethylene tubing would alleviate this issue and prevent cells from adhering to the inner walls of the device. Both options of 3D printing and machine manufacturing were explored, however it was determined that manufacturing the device would allow us to have direct control of the use of polytetrafluoroethylene for both the chambers and the transport tubing. In collaboration with a mechanical engineering student William Gill from the Schulich School of Engineering Machine Shop - a design was created and analyzed and produced from the desired materials.
Fluid Flow Analysis and Angle Optimization
In our initial design, most of the fluid would flow into the outside chambers from the PCR chamber. In order to make sure the fluid flows equally into the three chambers, fluid flow analysis was performed. On optimal angle for the tubings was found to be 20 degrees.