Team:HKIS/Entrepreneurship
Entrepreneurship
Background
The entrepreneurship page demonstrates that our kit will perform economically well considering market growth and other circumstances. Our team has worked tirelessly to ensure that our product is designed to meet consumer needs. After interviewing a member from the HKSAR seafood department and surveying many local fishers we have established a deeper understanding of what consumers are looking for in a detection system. The surveys and interviews eventually led us to design our product with storage, shipping, and instructional difficulties in mind, as well as leading us to utilize Lateral Flow Assays for clear visible results without requiring technical knowledge. This facet will include the target market, competing products, consumer opinions, and other resources.
Strong competitive edge: drastically reducing the cost of kit by >10 times
Our reagent cost is well optimized. While the original cas12a-RPA one pot detection protocol from Ding et al. utilizes 25μl reaction with 2.2μM of cas12a. We have done extensive optimization to lower the cost by >10X of the original protocal via buffer optimization (see wet lab section). We are able to achieve very high fluorescence signal with equal sensitivity (5 copies detection with naked eye). Such that we only take 10μl reaction with 0.5μM of cas12a, which is a 10X less reagent used provided that the most costly and consuming reagent is the crRNA transcription kit and cas12a protein. RPA mix is very cheap and only constitude <10% of the whole cost.
And hence, we have a huge competitive edge in terms of pricing. Where one cas12a (~200 USD) of 100μM could provide 400 reaction, and 1 crRNA transcrption reaction (~25 USD) and 1 RPA kit (1000 10μl reaction) reduces the cost of one reaction from 5 dollar from Ding et al to 0.5 dollar per reaction, which is a 10 fold reduction. Making it a very competitive price even to qPCR products which has similar costs. Moreover, a qPCR machine is not required with our eco-friendly kit.
Market Issue and Product Goal
What is the problem?
Our team has identified a significant issue with the current seafood industry, the oyster industry in particular. Globally, fishers do not have access to any diagnostic tools for determining if their produce is contaminated with Vibrio or Norovirus. The current products on the market for detection of Vibrio contamination are purely designed for usage in labs, and not as a product for aquaculturists and fishers.
SWOT Analysis
There are currently two existing categories of techniques for Vibrio detection, only one of which is commercially available. These two methods are qPCR amplification centered detection, and loop-mediated isothermal amplification (LAMP) centered detection. Both of these methods have massive drawbacks that prevent them from being realistic/viable options for the average fisher, and major companies that incorporate oysters into their products.
| RPA - Recombinase Polymerase Amplification (our solution) | |
|---|---|
| Strengths of the product: - High degrees of accuracy - can detect down to 5 copies of template |
Weaknesses of the product:
- Doesn’t display a spectrum of results (only positive/negative) |
| Opportunities: (for the application or commercial use of the product) - Applicable in small and large scale |
Threats: (to the viability or success of the product)
- Sample extraction and purification is technical - requires instructions |
| qPCR - Quantitative Polymerase Chain Reaction | |
|---|---|
| Strengths: - High degrees of accuracy |
Weaknesses:
- Takes longer to produce results (~45-60 mins) |
| Opportunities: - Widely known and used |
Threats:
- Portable, cheap detection systems |
| LAMP - Loop-mediated Isothermal Amplification | |
|---|---|
| Strengths: - High degrees of accuracy (specific) |
Weaknesses:
- Requires 6 core primers. |
| Opportunities: - Could be sent to developing countries (since it’s cheap and portable) |
Threats:
- Competing portable systems |
qPCR-based detection has several glaring issues. One of them is that the temperature conditions required for qPCR reactions are unachievable in an ordinary environment, and require an expensive thermocycling machine. Beyond this, machine handling requires an experienced technician and many technical instruments. The benefit of this method lies in the specific results that can be displayed. qPCR yields a spectrum result that can give more in-depth information than just a positive or negative result; the amplicons are also clean and usable for other assays, but that doesn’t concern the fishers using this product. Another massive downside to qPCR is the time it takes to yield a result. The entire qPCR reaction will take two to three days; in comparison, LAMP and RPA assays will take two arrays on average. This option is not viable for an average consumer.
LAMP on the other hand seems great in theory but the 61-68˚C activation temperature is difficult to achieve, and in that temperature most variants of Cas12a denature. Not only this, but LAMP reactions also require three pairs of primers per target, which are more complicated to design than RPA’s one set of primers per target and will require triple the primer quantities, thus driving up production costs. Beyond this, LAMP assays do not work with a lot of visualizing assays because end results tend to form dumbbell structures. LAMP assays also do not possess a distinct ssDNA phase, unlike our RPA assay, which means Cas12a must be designed with PAM sites in mind, further increasing primer design difficulty. This option is technically viable but has many disadvantages.
RPA takes the best of both worlds from qPCR and LAMP. RPA can operate at 37˚C and has a distinct ssDNA phase, meaning PAM site restrictions can be ignored in this specific reaction, increasing target site options and increasing the potential accuracy of our Assay. On top of that, the amplicons are produced in perfect condition, great for any other detection assays that require intact target sample DNA. RPA reactions are also very fast; qPCR reactions will take several days, but RPA assays will only take two hours to yield a result.
Business Plan: Costs/Profits/Resource plans
With the looming threat of COVID-19 still present, governments and global enterprises are rushing to design new and accurate methods of virus detection. Due to these developments, a stagnant market has shown increasing promise. The point of care/rapid detection market has been flourishing due to the application of many discoveries. Our team sees an opportunity to apply new technologies to the oyster agriculture industry, as the current mainstream Vibrio detection methodologies are inconvenient, highly technical, and expensive.
As of 2020, the international point of care diagnostics market is evaluated to be worth roughly USD 29.5 billion and is projected to reach USD 50.6 billion by 2025. The oyster market is also flourishing, estimated to be worth USD 8 billion as of 2019. In comparison, the movie market is estimated to be worth USD 17.4 billion. This shows that the oyster market is not small; in fact, it makes up 1/20th of the seafood market’s total evaluated value of USD 150 billion.
The oyster industry is a massive pool of wealth that our product will tap into, as essential oyster sauce-producing company Lee Kum Kee’s spokesperson has answered that they would definitely pay a premium for oysters that were tested and proven to be clean. However, due to how a key benefit of our kit is its universal applicability, and due to Vibrio’s prevalence in all other types of seafood, such as fish and mollusks, our team plans to extend our product to cover the seafood market as a whole.
Currently, though, our target is to raise the perceived oyster quality by certifying that oysters are uncontaminated at a local level. A large portion of the global oyster market lies in Asia, specifically Hong Kong, which has an incredibly active seafood market. Seafood imports and exports total around USD 2.55 billion annually. The Hong Kong oyster market is robust and is capable of investing in new technology for oyster contamination detection. Our product will also increase the range of oyster consumers, as results from our survey show that many potential consumers hold back from consuming shellfish due to fear of contamination. By proving products are uncontaminated, there will be a larger consumer base.
Target Consumers
To recap briefly on the target consumers mentioned on our implementation page, we plan to target seafood restaurants, oyster fishers/farmers, and supermarkets.
Consumer Interaction and Retail
Consumer interaction will be a challenge. We foresee our product being primarily retailed on online shopping sites such as Amazon, where we could receive product feedback from consumers on its product comments section. The target group most likely to purchase our product from Amazon would be large-scale oyster retail markets, high-end seafood restaurants, as well as large-scale oyster farms. But this does not fulfill the original mission our team set out to achieve, which is to improve Hong Kong’s local oyster production quality. Most local oyster fishers tend to not purchase products from online retailers, due to their rather traditional approach to oyster farming.
For us to tackle this problem, we have to find a better way to sell our product without the long-distance shipping and website navigation, which would turn away local fishers. Our solution is to make our products available in local fishing appliances retail stores, as there are many across Hong Kong in close proximity to oyster fishing hotspots.