Overview
We hope to manufacture a dengue virus detection kit which is convenient, fast, and effective. We expect that by detecting the dengue virus precisely, medical care and epidemic control of dengue could be undertaken at early stage, which could prevent dengue from developing into a severe and life-threatening situation.
Target Audience
- Proposed end users: Suspected dengue fever patients:
- Stakeholders:
-
Taiwan Centers for Disease Control (TCDC):
Airports, harbors
Imported cases are the main contributors to dengue epidemics in Taiwan. Patients who have been to a dengue affected foreign country and reported being bitten by mosquitoes may bring the virus back to Taiwan, which may result in a local outbreak thorough the local transmission.
The number of imported dengue cases in Taiwan is growing every year. In 2018, the total number of indigenous cases was 183, while imported cases numbered 350. In 2019, the imported cases increased to 540 while there were only 100 indigenous cases . The number of imported cases greatly surpasses the number of local cases and has become the main reason for Taiwan dengue outbreaks.
Thus, fever screening stations are set up by TCDC at airports and harbors. Everyone going through immigration will undergo infrared temperature detection. Anyone entering Taiwan who shows an abnormal temperature has to take a dengue rapid detection immediately. In Taiwan, about 70% of the cases of imported notifiable infectious diseases were dengue fever, and 40-50% of imported dengue fever cases were effectively detected by the fever screening.
The Local Department of Health:
Hospitals, clinics and health care centers
Based on the Regulations for Medical Devices set by the Ministry of Health and Welfare, a dengue virus detection kit is classified as a Class II medical device. It should be used under the supervision of professional medical workers. Thus, our product will be available to hospitals, clinics, and health care centers. The public can go to any medical institution and use our product for dengue diagnosis under the assistance of medical workers.
Currently nearly 2,000 medical institutions provide dengue fever rapid detection service. Those detection kits are controlled and distributed by the local department of health.
The Local Environmental Protection Bureau
Besides individual diagnostics, our product could also be used in vector surveillance in the environment. We could determine the vector infection rate in the environment by collecting and testing mosquitoes. Currently, nearly 3,350 residential areas conduct dengue vector surveillance every month under the auspices of the Environmental Protection Bureau. Thus, our product could be used to test the infection rate of vector at the same time.
Supply Chain
Goals
Short-term:
- Develop a detection kit focusing on dengue virus serotype 2.
- Sell the product to the relevant institutions.
Mid-term:
- Optimize our product.
- Develop a detection kit for all 4 serotypes of dengue virus.
- Establish the product in the Taiwan market.
Long-term:
- Look for an authorized distributor or authorized manufacturer to expand the foreign market and sell our product overseas.
- Focus on selling our product to developing countries and remote areas, where it has a comparative advantage.
How To Do
- Patent application: Class II medical device that uses peptides to detect the E protein of dengue virus particles.
- Quality test of the product:
- Functionality test in laboratory.
- Sensitivity: use whole virus particles (produced by a cooperating laboratory) to conduct the test.
- Specificity: use other flaviviruses which are in the same family as dengue virus (also obtained from a cooperating laboratory).
- Repeatability: conduct the test using multiple detection kits and different operators to ensure our product gets the same result.
- Use simulated blood samples (from a cooperating hospital) to repeat the functionality tests of the product.
- Use real blood samples (from a cooperating hospital) to conduct the functionality tests of the product, using double blind tests of patients’ blood samples and healthy blood samples.
- Test the product in comparison with current detection methods.
- Medical device license application:
Estimated time: 2 years. After producing the virus at the cooperating laboratory, we will conduct the first test in the first year and repeat the test a year later to ensure the stability of product. The tests will be done during the peak dengue infection period (summer).
Dengue virus detection kit is classified as a Class II medical device. It requires review and verification by Taiwan Food and Drug Administration (TFDA) to obtain a medical device license to be sold and used in Taiwan. Thus, our next step is to apply for medical device license from TFDA.
Value
Innovative
Using synthetic biology, we produce peptides utilized in a detection tool. The peptides have high flexibility. The peptides can distinguish different flaviviruses and serotypes of dengue virus, by changing their amino acid sequences. They can also be adapted to the mutations of the dengue virus.
Convenient
Our product can be used in the developing countries or remote areas, as no laboratory or instruments are needed for detection. The peptides have high stability, which makes them easier to be stored and transported to other countries.
Rapid
Our detection kit takes only 15 to 30 minutes to get results, which can help the appropriate units to take medical or preventive actions as soon as possible.
Domestic
Taiwan is currently using the detection kits imported from overseas. A domestic product could prompt Taiwan to build up its own biomedical industry chain, which can react immediately when outbreaks occur without relying on the foreign aid. This would help Taiwan to be more proactive in epidemic control.
Inexpensive
Peptides are easier to produce than antibodies, so the cost will be lower. The table below shows the differences between producing peptides and antibodies, and why peptides cost lower.
Difference between producing peptides and antibodies
Producing peptides using E. coli Generating antibodies using B cells Culture medium for E. coli cultivation is cheaper and is able to cultivate multiple varieties of E. coli. Culture medium for B cells is more expensive.
B cells cannot be produced on a large scale and may be contaminated.Induction of peptides can be done on a large scale through IPTG as lac operons appear in E. coli. Generation efficiency of antibodies is low, as antibodies are not main proteins produced by a B cell. There is no way to induce B cells to massively produce antibodies. Peptides can be purified efficiently as there are specific tags on the peptides. Purification of antibodies requires specific proteins (i.e. G protein), which also require time for production and purification. B cells have to be immortal before generating antibodies.
Challenges
It is impossible to get social approval in a short time after our product debuts as it requires time for users to use and approve it. Moreover, we have to compete with other current detection kits. The table below shows the comparisons between our product and other detection kits currently used in Taiwan.
As most of them have high accuracy, we have to ensure that our product performs better than them in order to persuade customers for using our product.
Users' Guide
In Vitro Diagnostics
Our product will be sold in boxes. There will be 10 detection kits and 10 lancets, separately packed in one box, with a 5 ml bottle of MES (2-(N-morpholino)ethanesulfonic acid) buffer. MES buffer is used to stabilize the pH of the blood sample and improve the lateral flow.
One detection kit, one lancet and one drop of MES buffer are used for one diagnosis.
- Use under the supervision of medical workers only. Wear gloves when using and wash hands after using.
- One bottle of MES buffer is sufficient for 10 detection kits. Other equipment should be properly disposed of after a single use.
- Store in a cool place away from direct sunlight.
- The validity of product is one year.
Vector Infection Rate Detection
A detection kit for use with mosquitos to test vector infection rate is sold in boxes without a lancet. Use one detection kit and MES buffer for each diagnosis.
- One bottle of MES buffer is sufficient for 10 detection kits. Other equipment should be properly disposed of after a single use.
- Store in a cool place away from direct sunlight.
- The validity of product is one year.
References
Dengue Fever Nationwide, Indigenous and Imported, Week1/2019-Week43/200〔Date of Onset 2018/12/30/2020/10/24〕, Taiwan National Infectious Disease Statistics System. https://nidss.cdc.gov.tw/en/nndss/disease?id=061
Shie-Liang Hsieh, Yu-Chih Lo, Pin Ling, Shu-Ying Wang, Chia-Yi Yu, Guey-Chuen Perng, …Chih-Peng Chang (2016). Dengue Fever in Taiwan: The New Perspectives from Epidemiology, Clinical to Fundamental Science, Ministry of Science and Technology. http://boe.tn.edu.tw/boe/wSite/public/Attachment/f1466038079289.pdf
Disease Surveillance Express, Taiwan Centers of Disease Control (TCDC) (2009-2020). https://www.cdc.gov.tw/En/Category/List/Nim1Frm1C1ELxEhJ_hdJBg
Hospitals With Dengue NS1 Rapid Test, Taiwan Centers of Disease Control (TCDC) (2020). https://data.cdc.gov.tw/en/dataset/dengue-ns1
Dengue Mosquito Investigation in Latest 12 months, Taiwan Centers of Disease Control (TCDC) (2020). https://data.cdc.gov.tw/en/dataset/dengue-mosquito-investigation-latest-12mM
The Serum Reagent of dengue virus Guidance (2017). http://regulation.cde.org.tw/data/downloadfile.php?sid=1269
Dengue NS1 Ag STRIP, Bio-Rad Laboratories (2020). https://www.bio-rad.com/webroot/web/pdf/inserts/CDG/en/70700_881097_EN.pdf
Pok KY, Lai YL, Sng J, Ng LC. Evaluation of nonstructural 1 antigen assays for the diagnosis and surveillance of dengue in Singapore. Vector Borne Zoonotic Dis. 2010;10(10):1009-1016. doi:10.1089/vbz.2008.0176
SD BIOLINE DENGUE NS1 AG TEST FOR EARLY CLINICAL PHASE OF DENGUE INFECTION, Standard Diagnostics Inc., Abbott (2020). https://www.globalpointofcare.abbott/en/product-details/sd-bioline-dengue-ns1-ag.html
SD product catalog, Standard Diagnostics Inc., Abbott (2020). ttps://ensur.invmed.com/ensur/contentAction.aspx?key=ensur.452313.S2R4E4A3.20181224.9265.4175105
SD product catalog, Standard Diagnostics Inc., Abbott (2020). https://ensur.invmed.com/ensur/contentAction.aspx?key=ensur.452313.S2R4E4A3.20181224.9265.4175105
DXID062018-EN-6 MULTISURE Dengue CE FA HR no bleed, MP biomedicals (2020)https://media.mpbio.com/productattachment/DXID062018-EN-6%20MULTISURE%20Dengue%20CE%20FA%20HR%20no%20bleed.pdf (include false rate)
ASIAGEN DENGUETEST KIT, Asiagen, (2020) https://asiagendenguetestkit.wordpress.com/principles-procedures/
Narayan Gyawali and Andrew W. Taylor-Robinson (July 26th 2017). Diagnosis of Dengue: Strengths and Limitations of Current Techniques and Prospects for Future Improvements, Dengue - Immunopathology and Control Strategies, Márcia Aparecida Sperança, IntechOpen, DOI: 10.5772/67680. Available from: https://www.intechopen.com/books/dengue-immunopathology-and-control-strategies/diagnosis-of-dengue-strengths-and-limitations-of-current-techniques-and-prospects-for-future-improve
© iGEM Team, 2020 CCU Taiwan
Patients present symptoms like fever, muscle and joint pain, and skin rash.
According to statistical data provided by Taiwan Centers for Disease Control (TCDC), from 2011 to 2019, the number of tests conducted for dengue diagnosis fell between 2,700 and 9,300. These numbers show the basic need for our detection kit. Based on the outbreaks in 2014 and 2015, the need for detection kits could reach 70,000 during a dengue outbreak.
By promotion of our product along with the spread of epidemics due to global warming, the demand for our product will increase continuously in the future.