Disorder Intro

Epidermolysis Bullosa (EB), often referred to as the “Butterfly Children” disease, encompasses a group of rare and severe inherited skin fragility disorders. It is caused by mutations in genes encoding structural proteins (e.g. keratins, collagens), which are crucial for maintaining skin integrity, particularly at the dermal-epidermal junction. The main types of EB include Epidermolysis Bullosa Simplex (EBS), Junctional EB (JEB), and Dystrophic EB (DEB). We have chosen this subtype due to its characteristically severe manifestations, the critical unmet need for accessible diagnostic tools, and the suitability of the causative mutations in the COL7A1 gene for detection via our integrated RPA and CRISPR-Cas12a platform.

Econ Pressure & Psychological Impact

• A European study (n=204) indicated that the total annual socioeconomic cost for patients with EB was approximately €31,390 per patient.
• European studies call for EB-specific subsidy systems, in Brazil, 28.85% of patients had malnutrition exacerbating quality-of-life decline.
• Brazilian study (n=31): 71% of patients had depressive symptoms (29% mild, 22.6% moderate-severe), far exceeding the general population rate (3.8%).

Traditional Prenatal Diagnostic Methods

• Amniocentesis is one of the more commonly used methods. The doctor uses ultrasound guidance to insert a needle through the abdominal wall into the amniotic cavity to extract a small amount of amniotic fluid (containing fetal exfoliated cells). However, this technique is an invasive procedure with a miscarriage risk.
• Chorionic Villus Sampling (CVS) offers the advantage of earlier detection timing. However, it remains an invasive procedure: the risk of miscarriage is slightly higher than with amniocentesis, and issues such as insufficient sampling or maternal cell contamination may also occur.
• Fetal Skin Biopsy was used in the prenatal diagnosis of EB in the past, especially for certain specific subtypes. Fetal skin tissue is obtained through ultrasound-guided puncture, and then histopathological examination and immunofluorescence staining are performed to observe the skin structure and protein expression to confirm the diagnosis. However, this method is highly traumatic and may cause irreversible damage to the fetus.

Advantages of Our Products and Technology

• This technique, based on cell-free fetal DNA (cffDNA) in maternal blood and combined with RPA isothermal amplification (Recombinase Polymerase Amplification) and CRISPR-Cas12a targeted cleavage, achieves ultra-early non-invasive screening at 8 weeks of gestation.
• One-step POCT workflow: From sample to result in less than 90 minutes (compared to weeks for sequencing), which is operable by minimally trained staff without reliance on specialized lab infrastructure.
• The cost is much lower than that of traditional methods, and this cost advantage facilitates large-scale population screening in low-resource settings.

This study focuses on the rare disease Epidermolysis Bullosa (EB), aiming to promote the prevention and treatment of rare diseases and enhance social awareness from multiple perspectives. The research concept is based on the survival plight of EB patients, with respect for the diversity of life as the core, and is committed to building a mechanism for the whole chain of “screening-diagnosis-treatment-guarantee”, and at the same time, prying open the long-term pattern of the fight against all rare diseases.

At the technical level, the test strip technology developed by the team is a non-invasive or minimally invasive way of sampling, which can be used to collect peripheral blood of pregnant women or saliva/blood of couples in early pregnancy, without complicated instruments, and is convenient for home use, and the process is simple and painless. The popularization of this technology can reduce the long-term consumption of medical resources caused by the birth of critically ill children, and will save resources to be invested in the broader field of rare disease prevention and treatment.

In terms of social impact, the research will focus on the “complete humanity” of rare disease groups through product publicity and dissemination of patients' stories, carry out public welfare activities to awaken society's attention to more than 7,000 rare disease groups, eliminate discrimination, and convey the concept of “preventable and controllable rare diseases”. We will focus on the “complete humanity” of the rare disease community, carry out public welfare activities to awaken the attention of society to the 7,000-plus rare disease communities, eliminate discrimination, convey the concept of “rare diseases are preventable and controllable,” promote the establishment of a sustainable environment for the survival and development of the patients, and help the human society to realize the collective goal of “eradicating rare diseases” and vision of sustainable development.

Our first product is a rapid, non-invasive lateral flow test strip for prenatal detection of Epidermolysis Bullosa causing mutations, specifically targeting pathogenic variants in the COL7A1 gene, as well as the high-frequency pathogenic variant in the LAMB3 gene, an in vitro detection platform called PrenatalEB-Detect.

This test strip, using the CRISPR-Cas12a system’s high specificity and sensitivity, combines isothermal amplification with CRISPR-based detection to pick up tiny amounts of fetal DNA from mom’s blood. It’s a game-changer for prenatal EB screening, especially in underserved areas, offering an affordable and easy-to-use early detection tool that helps folks make smart choices about their pregnancy.

Figure 1 The picture of our product

Our second product is a Fluorescence detection platform. This platform specifically recognizes disease-causing mutation sites by synthetic crRNA, combines with fluorescent reporter probes to achieve visual signal output, and completes the detection of disease-causing mutations in trace amounts of fetal free DNA (cfDNA), which has the advantages of being rapid, low-cost, and non-instrument-dependent.

Furthermore, compared to the popularity of our first product, our second product is more inclined to be used in hospitals and other professional medical institutions to make up for the inaccurate results brought about by improper handling of test strips, and these two products make up for each other's shortcomings, which greatly increases our competitiveness.

Fluorescence Detection Platform

Figure 2 Fluorescence Detection Platform

2.2.1 EB test strip

The EB test strip project targets two high-frequency disease-causing mutations in EB, COL7A1 and LAMB3, and designs PrenatalEB-Detect, an in vitro test platform based on the combination of isothermal amplification (RPA) and the CRISPR-Cas12a recognition system. has the following advantages comparing with existing assays:

1. Non-invasive:

Compared to traditional invasive tests such as amniocentesis, EB test strips provide a comfortable, needle-free testing experience with less physiological and psychological impact on the user.

2. Rapid Results:

EB test strips provide fast and reliable results, with results available in 125 minutes for timely decision making compared to days or weeks of waiting time for results from other traditional tests.

3. High-Sensitiity and Accuracy

EB test strips offer high accuracy and sensitivity for superior diagnostic performance and reliable results.

4. Cost-Effective

EB test strips are designed to be affordable, providing high quality at a low price compared to the $1,000 price tag of traditional testing methods such as amniocentesis, without compromising on performance.

2.2.2 Fluorescence detection platform

• It delivers exceptional detection sensitivity, enabling effective identification of low-abundance target nucleic acids. This allows for the reliable detection of trace pathogens or mutant genes in samples, such as viral nucleic acids at levels as low as a few copies per microliter in viral testing scenarios.
• Leveraging the CRISPR-Cas12A system’s specific recognition capability, the platform achieves superior specificity, accurately distinguishing subtle differences between target and non-target nucleic acids—including single-base mutations—thereby effectively avoiding cross-reactivity.
• With strong scalability, it can be adapted to diverse detection needs by designing different guide RNAs (gRNAs), enabling the detection of multiple pathogens, gene mutations, and other targets.
• It supports quantitative or semi-quantitative analysis of results through various signal detection methods (e.g., fluorescence, electrochemistry), facilitating more accurate determination of target content in samples and reducing human error.

In the world, the number of pregnancies in a year is estimated to be around 213 million, and just in China, there are more than 9 million babies born annually. Epidermolysis Bullosa (EB) is a life-threatening rare genetic skin condition, present in an estimated 20-45 per million live births. Although it is uncommon, the toll is tremendous—European studies estimate €31,352 (approximately $33,000), U.S. estimates run at $36,000 a year per patient—worldwide expenditures on EB-related healthcare are in the range of $200-400 million yearly for these patients alone. But even so, most cases of EB are still found outside the womb after birth. Virtually no screening methods are used during pregnancy, such as exome sequencing, which is costly, time-consuming, and inaccessible for broad population use.

Our products utilize CRISPR-Cas12a—a cost-effective, non-invasive, simple method to detect EB mutations in a short span of time (less than 1 hour). Modeled on the pregnancy test, it is composed of ready-to-use components. It can be used in maternal health clinics and primary health care facilities. The effective use of our products will promote early intervention, which in turn will help to lower the cases of congenital defects. This will significantly, in the long term, reduce the fiscal and medical burden on families and healthcare systems.

https://www.ncbi.nlm.nih.gov/books/NBK599531

https://www.stats.gov.cn/english/PressRelease/202402/t20240228_1947918.html

https://ojrd.biomedcentral.com/articles/10.1186/s13023-024-03328-1

https://pmc.ncbi.nlm.nih.gov/articles/PMC4727534/

In the following power-interest grid, we have mapped out stakeholder interest and power based on our goals of validating the feasibility of our prenatal EB screening platform, optimizing its design and implementation, and ultimately promoting its accessibility and clinical relevance.So, this analysis isn’t just about the biomedical innovation of CRISPR-Cas12a detection, but it also dives into the ethical, social, and commercial issues of introducing non-invasive prenatal testing for rare genetic skin disorders. We’ve already sorted and positioned stakeholders—like clinicians, researchers, biotech companies, patient advocacy groups, and regulators—based on their impact and involvement in our project. This strategic categorization helps us prioritize communication, guide collaboration throughout development, and efficiently allocate resources to each stakeholder.

Power-interest Grid

Figure 3 Power-interest Grid

This stakeholder map gives a clear picture of how different groups interact with our prenatal genetic screening platform. The stakeholder such as hospitals, clinical geneticists, and regulators are super involved and invested since they’re directly tied to medical use, patient safety, and approval processes. On the other hand, academic researchers, diagnostic equipment suppliers, and biotech investors, while important to innovation and funding, may show moderate influence or interest at the initial stages. Expecting parents and patient advocacy groups, though not decision-makers, are key end-users with growing interest as the product moves closer to application. This classification allows us to tailor our engagement strategies and focus our efforts on partnerships and communication pathways that will maximize clinical impact and accelerate responsible translation of our detection platform.

Figure 4 Stakeholder map

Our target customers include：

1. All persons planning to have a child and pregnant women in the course of pregnancy.

Demographics:

· Individuals or couples aged 25-40 that are planning to conceive or currently in the first or second trimester of pregnancy.

· Primarily located in urban and suburban areas, with medium to high health awareness and purchasing power.

Needs and PainPoints:

· Strong desire for children's health and early screening. (ACOG, 2020)

· Lack of awareness and access to some affordable early screenings.

· Concerns about invasive detection and expensive testing.

2. Medical Institutions in County-level or advanced areas.

Demographics:

·County-level hospitals, community clinics, and maternal health centers in developing or resource-limited regions, which lack innovative technology like genetic diagnostic tools due to cost and infrastructure barriers.

·Hospitals and other medical institutions in metropolitan areas with considerable funding for utilizing our Fluorescence detection platform.

Needs and Pain Points:

· There is a need for scalable, affordable screening tools that can be used by non-specialists.

· Limited budget and inability to provide advanced prenatal diagnosis. (World Health Organization, 2020)

· There is a need for a more accurate and acceptable Fluorescence Detection Platform used in advanced medical institutions.

3. Local Governments and Public Health Programs in Poverty-Stricken Areas

Demographics:

· Government health departments in rural, low-income, or minority regions, often supported by national maternal-child health initiatives.

Needs and Pain Points:

· High pressure to reduce the incidence of genetic birth defects.

· Budget constraints make high-end genetic testing unsustainable.

· Demand for proven, low-cost solutions that can scale across districts or provinces.

The price of the test strip : 99 RMB

According to our competitor research, current testing methods like NGS are expensive（3000-10000 RMB per product) and are not affordable to most people in China. Therefore, we aim to satisfy more people's needs and price our product at 99 RMB. Please refer to 6.2 Our Revenue and Cost Planning for details.

The price of the Fluorescence detection platform: 990,000 RMB

Because this product is positioned as a high-end, professional prenatal screening tool, we priced it at 990,000 RMB for advanced medical institutions.

The price of Detection Reagent Kit: 19,999

After purchasing our fluorescence detection platform and subsequent testing, customers will require the use of detection reagent kits. This reagent kit can meet the requirement of 100 tests.

We have developed different marketing programs for different target customers:

1.For expecting parents and family planners

Our goal is to provide individual and family programs for children through community health outreach, OB/GYN clinic partnerships, and digital platforms. By emphasizing ease of use, non-invasiveness, and affordability, we position our test suite as an empowering tool for informed decision-making. Educational activities at prenatal centers and social media content in local languages can greatly increase people's awareness and trust.

2. For primary healthcare providers in underserved areas and advanced areas

We will collaborate with local hospitals, rural clinics, and maternal health NGOs to introduce our screening kits as a practical, low-barrier diagnostic solution.Our training modules will ensure that even non-geneticists can test with confidence.By reducing reliance on expensive laboratory infrastructure, our products enable early detection without compromising efficiency in other areas. Furthermore, we will sell our Fluorescence Detection Platform to native advanced hospitals, offering them an additional way for patient to have a test.

3. For Government and Public Health groups

Our plan fits with the goal of lowering the frequency of birth defects in countries: to get public health and policy leaders involved. We also offer scalable, cost-effective solutions that are based on data on how things really work in the world. We market our products as high-impact, low-cost nstruments that promote equity in maternal and child health through pilot projects, bulk purchasing discounts, and public health campaigns that are co-branded with other organizations.

Furthermore, we will use popular online platforms like WeChat, Xiaohongshu, and JD Health to raise awareness through educational content, expert endorsements, and influencer collaborations. We will also make it easy to buy through official mini-programs and e-commerce stores. We want to establish trust and attract a lot of people to use our product by giving away trial kits, running interesting referral programs, and showing targeted digital ads to expectant parents, healthcare practitioners, and public health agencies.

Figure 5 PEST Analysis

Figure 6 Porter's five force Model

Figure 7 SWOT Model

However, the weakness of our EB Test Strip can be offset by our second product: the Fluorescence detection platform. Its professionalism and traits could increase users' trust and enhance their testing experience.

Thus, each of the shortcomings of our product can be effectively alleviated, helping us gain the competitive edge.

Figure 8 Business Canvas

Step 1: R&D(10 months):

Development in the lab, and carry out relevant scientific popularization events.

Step 2: Authority(14 months):

Experiment and wait for authorized permission. Product registration and approval.

Step 3: Marketing(18 months):

Setting up our sales channels on online shopping platforms like Taobao, JD, and Pinduoduo. First off, we’re aiming to grab 0.5% of the pregnant women market in China. Plus, we’ll team up with medical institutions for wholesale marketing. We’re hoping to partner with 5 medical facilities in the first year in 5 metropolitan cities after the product launch.

Step 4: strength(32 months):

We're gonna keep expanding our market, ramping up our marketing efforts, and hiring top-notch salespeople. The goal is to increase our market share by 10% every year. We'll boost our brand awareness by promoting and advertising on well-known media so that more people know about our products. And we'll keep on researching and developing to improve the product quality and service, so as to solidify our product's market position.

Figure 9 Our Business Step and Milestone

For our Test Strip Product, we aim to gain a competitive edge with an accessible two-digit RMB price (99 RMB), addressing the affordability barrier of traditional Epidermolysis Bullosa (EB) screening methods like amniocentesis, which cost 2,000–7,000 RMB and remain unaffordable for many Chinese families. Based on the National Bureau of Statistics of China (2024) data showing 9.02 million new births in 2024, we target capturing a 5 per 1,000 market share in our first year, projecting sales of approximately 45,100 test strips and estimated revenue of 4,464,900 RMB.

Our fluorescence detection platform mainly consists of a fluorescent - labeled enzyme and a detection reagent kit. Once purchased, the fluorescence detection platform can be used for a long time, and there is no need for additional purchase costs later. According to our research on similar products on the market, the selling prices of fluorescence microplate readers fall into different tiers. Low - end products cost approximately 20,000 - 50,000 RMB. Mid - range products range from 100,000 - 245,000 RMB. High - end products are about 340,000 RMB. We believe that our product should meet the needs of most people, so we position our fluorescence microplate reader as high-end , with the cost taking the median value of 172,500 RMB.Our fluorescence detection platform can target multiple gene mutation diseases, compared to traditional detection of single mutation diseases. So our product range is more extensive, priced at 990,000 Yuan.

The fluorescence detection kit includes consumables such as fluorescent probes, cutting enzymes, and guide RNA required for detecting the target gene, as shown in the figure below. After we consulted the R & D personnel, the relevant composition is a trade secret and has not been disclosed yet, so we have obscured some content. This composition can be used to produce reagent kits for approximately 100 tests in total. The total cost is 9,673.3 RMB. So we priced it at 199 yuan, which can meet the needs of most hospitals by changing the price.

Figure 10 The Cost of Raw Material Composition

Estimated land cost

Estimated amount: 375000 RMB

Assumption basis: we choose to build our own factory in Jiangsu Yancheng Biological Park, the plant covers an area of 1000 square meters, the government in the original price of 450,000-630,000rmb have a 70% discount subsidies, after the discount price of 315,000-435,000rmb in Yancheng Economic and Technological Development Zone or the South China Sea Future City Biomedical Industrial Park, to enjoy the tax relief and infrastructure support.

Estimated cost of factory construction

Estimated amount: 2250000 RMB

Assumption basis: biosynthesis factory construction through the calculation of the minimum 1500000rmb maximum 3000000rmb The cost includes construction costs, clean room, etc., with reference to the 2022 Yancheng, a medical device factory total cost of 2800000rmb and the project scale is close to

Equipment cost estimation

Estimated amount: 3,500,000 RMB

Assumption basis: 350w of the cost is the cost of equipment, including fermentation equipment, bioreactors, purification equipment, testing instruments, etc., the expected cost range of 2,000,000-5,000,000rmb. If the choice of equipment needs to be balanced with the degree of automation, such as semi-automatic production line can save 1,000,000rmb, but the cost of labor increase ) Used equipment can reduce the cost of 30% -50%, but need to verify the performance (such as through the “experimental easy to purchase” platform procurement).

R&D cost estimation

Estimated amount: 5,000,000 RMB

Assumption basis: We expect that R&D will require CRISPR reagents, experiments, clinical validation, as well as patents and certified domestic invention patents (1-2 items). This cost we have overestimated, granting a budget of 5,000,000 RMB.

Other cost estimates

Estimate: 750,000 RMB

Assumption basis: other cost data provided by other synthetic biology and medical device R&D factories, this cost includes EIA and approvals, design, installation, commissioning, initial raw material procurement and pilot production, staff training and operation startup, estimated around 500,000-1000000rmb

Annual Fixed Costs

Labor：2340000 RMB

Given that we will build our factory in Yancheng and the average revenue of native workers who work in private companies is 78114 RMB (National Bureau of Statistics of China, 2024), our company is prepared to offer employees an annual salary of 78000 RMB and we will hire 30 workers.

Employee Welfare: 500000 RMB

We want to offer workers some treatments, like year-end and business bonus.

Power: 600000 RMB

According to our scale and construction of our factory, our power cost is nearly 600000 RMB every year.

Market Promotion:850000 RMB

Our company is prepared to conduct our promotion strategies：

1.Content production and educational promotion

We will spend 225000 RMB producing professional videos, online lectures, and distributing popular science articles.

2.Social media and platform advertising

300000 RMB need to be spent to advertise on WeChat, Xiaohongshu, TikTok and other online platforms.

3.Trial packs and promotional samples

Producing free or low-priced trial packs and sending them to target users and primary medical institutions cost 27500 RMB.

4.Operation of online shopping malls and mini-programs

100000 RMB for maintaining customer service, user interaction and conversion support for e-commerce platforms (such as WeChat Mini Programs and JD Health)

Variable Cost

Cost Per Product:15 RMB Total:676500 RMB

Including the cost of the test strip, degradable box, and decoration on the box. According to our research, some genetic materials (1200 RMB per unit) and one test strip (1300 RMB per unit) can produce

100-200 units product; other materials cost little, so we estimate that the cost of every product per unit is 15 RMB.

Supply Chain Per Product:3 RMB Total:135300 RMB

Including the cost of distribution and delivery charge.

As we explained above in revenue part, Our fluorescence detection platform mainly consists of a fluorescent - labeled enzyme and a detection reagent kit. Once purchased, the fluorescence detection platform can be used for a long time, and there is no need for additional purchase costs later. According to our research on similar products on the market, the selling prices of fluorescence microplate readers fall into different tiers. Low - end products cost approximately 20,000 - 50,000 RMB. Mid - range products range from 100,000 - 245,000 RMB. High - end products are about 340,000 RMB. We believe that our product should meet the needs of most people, so we position our fluorescence microplate reader as a mid - range one, with the cost taking the median value of 172,500 RMB.

The fluorescence detection kit includes consumables such as fluorescent probes, cutting enzymes, and guide RNA required for detecting the target gene. After we consulted the R & D personnel, the relevant composition is a trade secret and has not been disclosed yet, so we have obscured some content. This composition can be used to produce reagent kits for approximately 100 tests in total. The total cost is 9,673.3 RMB.

Figure 11 Finance Forecast