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+ | <h1 align="center">Design</h1> | ||
+ | <br> | ||
+ | <p align="center">To design criteria for a lateral flow assay, we developed an outline of important design specifications determined via a simple market analysis. </p> | ||
+ | <br> | ||
+ | <br> | ||
+ | <center> | ||
+ | <button class="btn-link" data-balloon-length="large" data-balloon="A performance time of 30 min or less would be optimal based on the World Health Organization’s current ASSURED criteria for rapid tests. Current tests on the market produce results in 10-30 min with most producing a result within ~15 min. As this is the market standard, 15 min was set as the design criterion." data-balloon-pos="up"> | ||
+ | <a class="foo" href="#Time" > <img src="https://static.igem.org/mediawiki/2017/c/c0/T--Duke--DesignTimeButtonBefore.png" height="190"/> <img src="https://static.igem.org/mediawiki/2017/6/67/T--Duke--DesignTimeButtonAfter.png" height="190" /> </a> | ||
+ | </button> | ||
+ | <button class="btn-link" data-balloon-length="large" data-balloon="Current RDTs test for an immune response. However, this type of test requires the body to undergo seroconversion. This amounts to a minimum of a 4-week delay in the case of HIV; exact timing of seroconversion varies between persons." data-balloon-pos="up"> | ||
+ | <a class="foo" href="#DetectionWindow" > <img src="https://static.igem.org/mediawiki/2017/7/7c/T--Duke--DesignWindowButtonBefore.png" height="190"/> <img src="https://static.igem.org/mediawiki/2017/a/a2/T--Duke--DesignWindowButtonAfter.png" height="190" /> </a> | ||
+ | </button> | ||
+ | <button class="btn-link" data-balloon-length="large" data-balloon="The third metric considered the cost associated with testing. In Rwanda, the average cost of testing positive for a HIV is $1300+ USD. Considering that the average individual wage is ~$0.52 USD, an entire family must work 40 hour per person per week just to pay for the test. Therefore, our test should be deliverable for under $1USD. " data-balloon-pos="up"> | ||
+ | <a class="foo" href="#Cost" > <img src="https://static.igem.org/mediawiki/2017/thumb/3/34/T--Duke--DesignCostButtonBefore.png/600px-T--Duke--DesignCostButtonBefore.png" height="190"/> <img src="https://static.igem.org/mediawiki/2017/thumb/a/a5/T--Duke--DesignCostButtonAfter.png/600px-T--Duke--DesignCostButtonAfter.png" height="190" /> </a> | ||
+ | </button> | ||
− | < | + | <button class="btn-link" data-balloon-length="large" data-balloon="The lateral flow assay we developed could be expanded past HIV and ZIKV to test for other enveloped viruses by changing only the antibody used in the assay. " data-balloon-pos="up"> |
− | < | + | <a class="foo" href="#Modularity" > <img src="https://static.igem.org/mediawiki/2017/thumb/d/da/T--Duke--DesignModularButtonBefore.png/600px-T--Duke--DesignModularButtonBefore.png" height="190"/> <img src="https://static.igem.org/mediawiki/2017/thumb/8/83/T--Duke--DesignModularButtonAfter.png/600px-T--Duke--DesignModularButtonAfter.png" height="190" /> </a> |
− | < | + | </button> |
− | + | </center> <br> | |
− | </ | + | |
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− | < | + | <button class="btn-link" data-balloon-length="large" data-balloon="According to the WHO guidelines, the sensitivity and sensitivity of RDTs for HIV is ~50 virions/mL or 150 copies of RNA/mL and the specificity should be ~98-99%. This set our final design criteria. " data-balloon-pos="up"> |
− | + | <a class="foo" href="#Modularity" > <img src="https://static.igem.org/mediawiki/2017/thumb/d/d8/T--Duke--DesignSensitivityButtonBefore.png/600px-T--Duke--DesignSensitivityButtonBefore.png" height="190"/> <img src="https://static.igem.org/mediawiki/2017/9/96/T--Duke--DesignSensitivityButtonAfter.png" height="190" /> </a> | |
− | </ | + | </button> |
+ | |||
+ | <button class="btn-link" data-balloon-length="large" data-balloon="Furthermore, a lateral flow assay proves to be a natural choice for its ease of use and familiarity to the target population. Therefore, using the generalized lectin-antibody combo with the lateral flow assay would allow for a RDT capable of testing for viral infection quickly and accurately." data-balloon-pos="up"> | ||
+ | <a class="foo" href="#Modularity" > <img src="https://static.igem.org/mediawiki/2017/thumb/0/0b/T--Duke--DesignTechButtonBefore.png/600px-T--Duke--DesignTechButtonBefore.png" height="190"/> <img src="https://static.igem.org/mediawiki/2017/thumb/1/1a/T--Duke--DesignTechButtonAfter.png/600px-T--Duke--DesignTechButtonAfter.png" height="190" /> </a> | ||
+ | </button> | ||
+ | |||
+ | <button class="btn-link" data-balloon-length="large" data-balloon="Our design utilized the lectin, griffithsin (GRFT). GRFT binds to glycosylated sugar chains that are found on the envelop of some viruses including HIV. The GRFT binds to HIV glycoprotein gp120, which is highly conserved across variants of HIV. Thus the final RDT developed would be able to detect HIV irrespective to variant and subtype. Therefore using this lectin to create a lateral flow assay would result in a test superior to existing options" data-balloon-pos="up"> | ||
+ | <a class="foo" href="#Modularity" > <img src="https://static.igem.org/mediawiki/2017/thumb/7/7d/T--Duke--DesignGRFTButtonBefore.png/600px-T--Duke--DesignGRFTButtonBefore.png" height="190"/> <img src="https://static.igem.org/mediawiki/2017/a/a5/T--Duke--DesignGRFTButtonAfter.png" height="190" /> </a> | ||
+ | </button> | ||
+ | <br> <br> <br> | ||
+ | <center><p> | ||
+ | With these design criteria in mind, we developed the following lateral flow assay. | ||
+ | <br> <br><br> <h1> Solution: Lateral Flow Assay </h1> | ||
+ | </p> </center> | ||
+ | <br> <br> <br> <br> <br> <br> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/8/8a/T--Duke--LFA_DesignTry3.jpg" width="900"> | ||
+ | </center> | ||
+ | </center> | ||
+ | <p> In order to achieve this lateral flow assay our team focused on engineering a thermostable variant of griffithsin which would be needed to create the assay and allow the diagnostic to have a longer shelf life. If this part were to be produced, then the rest of the assay would only require an assembly of the required parts. The major focus of our wet lab was focused on developing this thermostable variant of griffithsin.</p> | ||
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Latest revision as of 03:06, 2 November 2017
Design
To design criteria for a lateral flow assay, we developed an outline of important design specifications determined via a simple market analysis.
With these design criteria in mind, we developed the following lateral flow assay.
Solution: Lateral Flow Assay
In order to achieve this lateral flow assay our team focused on engineering a thermostable variant of griffithsin which would be needed to create the assay and allow the diagnostic to have a longer shelf life. If this part were to be produced, then the rest of the assay would only require an assembly of the required parts. The major focus of our wet lab was focused on developing this thermostable variant of griffithsin.