Team:CCU Taiwan/Poster
DENDETX
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Project Goals
- Develop a rapid test for dengue fever to improve treatment outcomes.
- Develop a method to mass-produce peptides for virus recognition, reducing cost.
- Raise awareness of dengue fever in scientifically illiterate and high-risk groups.
Introduction
Dengue fever occurs in tropical and subtropical regions. Half of the world population is now at risk, with 390 million infections every year, causing dengue fever to be listed by WHO as a top 10 health threat.
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Dengue fever can progress into more serious conditions like dengue hemorrhagic fever (DHF), which has a 20% mortality rate. If conditions continue to worsen, patients experience dengue shock syndrome (DSS), which has a mortality rate up to 40%. An estimated 500,000 patients develop DHF or DSS, resulting in about 25,000 deaths every year.
Taiwan is in the area under the threat of dengue virus. From 2014 to 2015, there were over 44,000 confirmed cases during a dengue fever outbreak that spread from Tainan to other cities, causing 228 deaths. Recently, there was local transmission of dengue fever in Taoyuan, showing that the potentially affected population keeps increasing.
There is no specific treatment or vaccine for dengue, and health care systems are still facing a challenge from dengue fever.
Dengue fever can progress into more serious conditions like dengue hemorrhagic fever (DHF), which has a 20% mortality rate. If conditions continue to worsen, patients experience dengue shock syndrome (DSS), which has a mortality rate up to 40%. An estimated 500,000 patients develop DHF or DSS, resulting in about 25,000 deaths every year.
Taiwan is in the area under the threat of dengue virus. From 2014 to 2015, there were over 44,000 confirmed cases during a dengue fever outbreak that spread from Tainan to other cities, causing 228 deaths. Recently, there was local transmission of dengue fever in Taoyuan, showing that the potentially affected population keeps increasing.
There is no specific treatment or vaccine for dengue, and health care systems are still facing a challenge from dengue fever.
Inspiration: Interaction between the CLEC5A and E protein suggest proteins might be effective in detection
After infection by the dengue virus, a nonstructural protein, NS1, appears in blood serum early in the infection. The current method of detecting dengue fever uses antibodies to recognize NS1 antigens. Nevertheless, the cost of the antibodies is high, the antibodies are hard to purify, and the procedure of making antibodies is time-consuming.
We took a step back to seek an alternative. We aimed to find something smaller and easier to produce to detect dengue virus instead of using antibodies. Symptoms of DHF and DSS result from interaction between C-type lectin domain, family 5, member A (CLEC5A) on macrophages and the envelope protein (E proteins) of dengue virus.
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We found the sequences from CLEC5A most likely to bind the dengue virus and confirmed this using Rosetta simulations. We believe these peptides have potential to detect the dengue virus.
We designed a detection kit with these peptides that can bind to the E protein of dengue virus. This would allow people who may have dengue fever or live in an outbreak region of dengue fever to get treatment or protection and control the spread early.
We took a step back to seek an alternative. We aimed to find something smaller and easier to produce to detect dengue virus instead of using antibodies. Symptoms of DHF and DSS result from interaction between C-type lectin domain, family 5, member A (CLEC5A) on macrophages and the envelope protein (E proteins) of dengue virus.
We found the sequences from CLEC5A most likely to bind the dengue virus and confirmed this using Rosetta simulations. We believe these peptides have potential to detect the dengue virus.
We designed a detection kit with these peptides that can bind to the E protein of dengue virus. This would allow people who may have dengue fever or live in an outbreak region of dengue fever to get treatment or protection and control the spread early.
Linear Array Epitope
Linear array epitope (LAE) is a technique to produce tandem-repeated sequences (TRSs). After the procedure, the TRSs would be expressed as a long-chain peptide which can be cleaved by an enzyme, resulting in many identical peptides.
The procedure of LAE:
For the primer design, TR-PCR primers need to be partially complementary to each other and AD-PCR primers need to have restriction sites.
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In TR-PCR, those primers act as both primer and template and they are partially complementary. The TRSs can be produced during the PCR cycling process. In AD-PCR, restriction sites need be introduced into the products of TR-PCR by adding the AD-PCR primers in PCR.![]()
The procedure of LAE:
- Design DNA oligo primers.
- Template-repeated PCR (TR-PCR).
- Adaptor PCR (AD-PCR).
For the primer design, TR-PCR primers need to be partially complementary to each other and AD-PCR primers need to have restriction sites.
In TR-PCR, those primers act as both primer and template and they are partially complementary. The TRSs can be produced during the PCR cycling process. In AD-PCR, restriction sites need be introduced into the products of TR-PCR by adding the AD-PCR primers in PCR.
Parts
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Modeling: Interaction between the tandem-repeated sequence peptide and E protein
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The detection kit - DENDETX
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Results
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Human Practices
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Inclusion
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References
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