About This Project
M. tuberculosis is slow growing pathogen, TB in early infection is difficult to diagnose correctly, so culture is the WHO-recommended gold standard for TB diagnosis, but it may take more than 1 to 3 months. The patient may inadvertently become a spreader while waiting for the results. We propose a solution using Raman spectroscopy that can detect pathogen more accurate than the current test. If our results are positive, we should be able to early detect and treat TB to prevent spread of disease.
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What is the context of this research?
In recent years, the application of Surface enhanced Raman scattering (SERS) as a detection platform is more popular1 2, SERS so that the weak Raman signal enhanced 1011 times. we proposal the strategy consists of two phases: (i) Antibody or aptamer conjugated nanoparticles as probe, that can capture and concentrate the pathogen protein form the sputum sample, antibody can concentrate and confirm the pathogen specificity, and (ii) Raman spectrometer scanning Raman tag on nanoparticle surface. SERS technology can amplify Raman tag signal to 1011 times. Therefore, we are able to detect TB in the pathogen is very rare period. By developing the Raman-based system to detect a particular target protein, we can potentially adapt this technology to diagnose a particular disease.
What is the significance of this project?
M. tuberculosis grows slowly. In the early stages of infection, the number of bacteria is very small so it is difficult to diagnose the disease. Traditional diagnosis (Smears Acid-Fast bacillus staining) needs 1 to 3 months to culture and is very time consuming. As a result, TB in early infection is difficult to diagnose correctly. We are proposing may be able to accurately detect tuberculosis early in infection. If the results are successful, it can become an important tool for prevention and treatment of tuberculosis. Our design provides a cheap, sensitive, accurate, and rapid diagnostic tool that can effectively prevent the spread of tuberculosis. Additionally, our detection model can theoretically be adapted to detect any particular target, allowing expansion to detect other diseases.
What are the goals of the project?
After infection, M. tuberculosis will secrete many proteins into the sputum and blood, such as Ag85b, ESAT-6, CFP-10 and so on. In early infection, the bacteria is rare, but it still will secrete a lot of ag85b protein into the sputum and blood, so our method is to use an anti-ag85b antibody conjugated Fe2O3 nanoparticles, and then another anti-Ag85b aptamer conjugated Au NP, which also conjugated DTNB. We plan to add the antibody–Fe NP and aptamer–Au NP-DTNB into sputum , after the magnet aggregates and then use the spectrometer to detect the Raman signal of the DTNB. We combine specificity of antibodies and sensitivity of SERS . The model can be applied to any antigen, thus making a detection system that will work for a multitude of diseases.
We count on your support to acquire the materials necessary to run experiments and make progress on our project. We rely entirely on donations and contributions from sources such as payout by ourselves and fundraising opportunities like this. Although we can rent Raman spectrometer, but the rental costs of Raman spectrometer is about 16 US Dollar per sample, we need for a large number of tests in the research process. And we need to modify the sample holder according to the different samples pretreatment, but rented instruments cannot be modified. And, we will also develop other research in the future, so we need to spend money to build a spectrometer platform for future research. The SERS technique is based on nanoparticles, and different sizes of nanoparticles will directly affect the effect of SERS, so we will synthesize different sizes of nanoparticles for testing. and we will buy specific antibodies. And self-screening and testing of aptamers.
1. this project will begin on November 30, 2017, At the beginning we will produce ag85b protein for screening anti-ag85b aptamers. in the same time, Construction Raman Spectral System. It will finish before Jan 31, 2018.
2. Synthesis Fe2O3 nanoparticles and Au nanoparticles before Mar 31, 2018.
3. Antibody and aptamer are modified on the nanoparticle before jun 30, 2018.
4. Ag85b detection test before Nov 30, 2018.
Oct 09, 2017
Jan 31, 2018
Construction Raman Spectral System
Mar 31, 2018
Fe2O3 Nanoparticles synthesis and conjugated with antibody
Jun 30, 2018
Au Nanoparticles synthesis and conjugated with aptamer and DTNB
Nov 30, 2018
ag85b protein detection test
Meet the Team
We are from different scientific fields of researchers, the current members of a total of five, respectively, from the Biomedical Science, Nanotechnology and Chemistry. We are working in different institutions, but we are interested in development of detection platform. And Jiunyi Shaw has a startup company, the startup company's space as our research base to share each other's expertise and experience. We continue to find partners that have same belief, to carry out the detection system.
Jiunyi Shaw graduated from the Institute of Molecular Medicine at National Cheng Kung University. He majored in Molecular Biology and Cell Biology in college. Jiunyi worked as a Senior Researcher at the Institute of Biomedical Sciences of the Academia Sinica for 4 years, and then worked at Cancer Molecular Diagnostics Laboratory of Chang Gung Medical Foundation for 4 years. He is currently performing aptamer development at Gene Z Biotech Co., Ltd.
I graduated from the Institute of Oral Medicine at National Cheng Kung University. I earned my Master's in Nanobiotechnology. In the past , I worked in the RNAi core facility and genomic core facility in the Academia Sinica. Now I am working on new drug research.
I earned my Master's degree in Molecular Medicine at National Cheng Kung University in Virology. After that I became a research assistant at Academia Sinica for a few years and joined the Ph.D. program. Now I am a Ph.D. candidate.
My research interests cover from basic research to clinical applications. Currently, my project focuses on how to detect the pathogens in the important diseases.
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