2.  The HIV virus can mutate in HIV positive patients taking Anti-Retroviral Therapy (ART)  Their HIV strain has now become drug resistant (DR), and their current medication no longer works  This increases the patient’s viral load, and creates the possibility of passing on a drug resistant strain of HIV

3.  To create a device which can qualitatively detect if an HIV positive patient’s ART is working or if their strain has mutated

4.  Create an automated platform which combines RNA extraction, a PCR Thermocycler, and Gel Electrophoresis to give a qualitative result.

5.  Use RNA extraction methods to purify RNA  Send a single strand of RNA to PCR group  PCR group turns single strand of RNA to cDNA (complementary DNA) via RNA transcriptase.  Then use PCR to amplify DNA segment  PCR group sends amplified DNA to electrophoresis group

6.  Electrophoresis group runs samples through gel, sample can then be analyzed.  If a band is present on a specific segment of the gel, then HIV is present.  Can conclude the patient’s HIV strain is drug resistant, due to the presence of HIV

7.  Steps of RNA extraction:  Lyse serum  Adjust binding conditions  Bind to glass mesh  Remove contaminants Inhibitor Removal Buffer Wash Buffer  Elute

8. Roche : High Pure Viral RNA Kit

10.  Obtain sample from RNA team  Convert RNA to cDNA  Amplify DNA via PCR  Heat and cool sample according to protocol  Export to Gel Electrophoresis team

12.  Make Agarose Gel with Ethidium Bromide  Load amplified DNA from PCR/Thermocycler into gel  Run voltage (100V) through gel for approximately 20-40 minutes  View gel under UV light to determine presence of HIV  Visible due to ethidium bromide tag

13.  Building our own box based on open source designs  Looking for inexpensive, safer alternatives to ethidium bromide  Create safe waste disposal protocols if ethidium bromide must be used.

14.  Designing for Developing Nations.  This means Resource Limited Settings  Size should be reasonable  If it is too large it will be too expensive to ship  Cost  Must be incredibly reliable  Overly complex designs are more likely to break  Would like to be able to run between 20-100 patient samples in a day  Blood pooling methods