Difference between revisions of "Team:CSMU Taiwan/Demonstrate"

Overview

In addition to comply to all rules and policies approved by the iGEM Safety Committee (please see our Safety page), we self-evaluate the safety of our project with the Risk Mitigation Plan referring the selective agent and dual use research management regulations of TCDC.

This year, we design an aptamer-type rapid diagnostic kit called influsensor, trying to solve the disadvantages of screening on the influenza nowadays. (please see our Description page)

There are two main products in our project:

1. The aptamer with high affinity and specificity designed for rapid diagnostic test
2. Influensor (aptamer-type rapid diagnostic kit) that is extremely easy for doctors to use

Overall, our the aptamers selected out by the target proteins NPA BBa_K2951004 and HA3 BBa_K2951014 have been proved to possess the properties of high affinity and specificity through our experiments, indicateing that there is a potential to replacing the current antibody-type rapid influenza diagnostic strip. (please see our Entrepreneurship page)

Aptamers

This year, we acquire four successful experimental results related to produce aptamers with high affinity and specificity in our project:

Target protein with optimized sequence

The production of the target protein is the preprocess of SELEX. However, our original yield of NPA protein and NPB protein is too low to supply the need for SELEX. We optimized the sequence and compare the absorbance standard curve before and after the sequence optimization.

The fugures indicate that the protein yield of both NPA protein and NPB protein significantly increased after the sequence optimization.

SELEX

In order to acquire the aptamers of target proteins, we conducted the SELEX experiment. After 6 rounds of SELEX for every protein respectively, the aptamers of each target proteins were obtained. The PDF files below are the sequenced results of the aptamers:

• NPA PDF
• NPB  PDF
• HA1  PDF
• HA3  PDF

ELISA

After conducting the SELEX experiment. Using ELISA to confirm the titer value of the aptamer towards target protein enables to test two important characters we intend to understand about the selected aptamers: the affinity and the specificity.

On the figure of NPA(200X)-NPA, all of the aptamer of NPA protein tested in affinity test have significantly dose-dependent property.As a result , all of the aptamer have high affinity toward the NPA protein. On the figure of NPA(200X)-NPB, all of the aptamer of NPB protein tested in specificity test have significantly lower value comparing with the test of NPA protein. Then, we found that the NPA-4 aptamer and NPA-5 aptamer have the higher affinity toward NPA protein and lower affinity toward NPB, which means NPA-4 aptamer and NPA-5 aptamer have higher specificity. As a result, we chose NPA-4 aptamer and NPA-5 aptamer to experiment in the next stage.

On the figure of ApHA3-4, there is a significant logarithmic relevance between HA3-4 aptamer and the target protein HA3, indicating the the high affinity of HA3-4 aptamer. In addition, there is a lower relevance between the aptamer and the non-target protein, indicating the the high specifity of HA3-4 aptamer. Also, there is a lower value when comparing with the target protein. As a result, we found that HA3-4 aptamer can effectively distinguish the two protein.

Rapid diagnostic test strip

(Please see our Results page)

Influsensor

Strip

Casing

Swab

Influsensor is an aptamer-type rapid diagnostic kit with the characteristics of low cost, fast production and development, high stability, low false-negative rate, and animal free. (Please see our Design page)

Influsensor is made of aptamer-type rapid diagnostic strip, casing, and swab. We design every component of the kit with ingenuity. We also demonstrate the user manual and the video of user guide. All of our designs are aim for making doctors manipulate the kit easily. (Please see our Design page)

User manual

User guide

Because of all the advantages and features of the aptamer, we integrate all the benefits of atpamers and design a model containing all the aspects of a disease: Rapid screening, Prediction, Prevention, and Treatment. The model is called CSMU_Taiwan Influsystem.

(Please see our Description page and Design page)