Team:ASIJ Tokyo/Implementation


When we first proposed to construct a detection kit for breast cancer, we wanted to make sure that our detection kit would be implementable in the real-world market. Therefore, we used a combination of our human practice interviews and consultation with entrepreneurs to help shape and influence our project design.

Using Aptamers and AuNPs as Detection Component

To develop a convenient test kit, we first performed research on different ways to help detect our biomarkers. Through our literature review, we discovered the prominence of aptamers, which are short oligonucleotides that can bind to a specific target, in this case, our biomarker. Aptamers have been compared to antibodies in their ability to specifically bind to target molecules, and are significantly cheaper and convenient compared to conventional antibodies (McKeague, 2017).
With our discovery of aptamers, we then thought of methods of utilizing aptamers to help detect our biomarkers in a cell-free environment. We came across methods like using ELISA and measuring fluorescence intensity, however, many of these methods are not portable and requires the use of expensive experiment. For example, not only does ELISA require the use of a 96-well plate reader to measure for absorbance, but it also requires the use of antibodies which defeats the purpose of utilizing aptamers. These limitations led us to use gold nanoparticles, which are versatile devices for detection based on their aggregate and non-aggregate modes. When gold nanoparticles undergo a change in mode, this results in a change in the color of the nanoparticle solution. Therefore, we hypothesized that by monitoring the color change of our AuNP assay, we will be able to detect our biomarkers using aptamers.
Using colorimetric detection has many benefits that can aid with the convenience and implementation of our detection device. Our AuNP assay can be constructed by just incubating the aptamers with the AuNP overnight, before adding the biomarker of interest. Furthermore, we can eliminate the use of specified equipment from laboratories by combining our AuNPs with lateral flow immunoassay strips, which allows for a portable and low-cost detection system (Pan et al., 2018). We plan on developing software next year that can help quantify the concentration of biomarkers based on the color of the AuNP solution, which will help test for and indicate signs of breast cancer.
Figure 1: Four-Step Approach to Detection Kit Implementation(Aptamers, Gold Nanoparticles, Lateral Flow Test, Analysis Software)
Created with

Sensitivity and Specificity

One of the largest concerns for our project was the issue of false positives and inaccurate diagnostic results. With breast cancer stigma being a huge issue in Japan, we wanted to make sure our detection kit followed local and global guidelines to create a convenient device for women around the world.
While we have not been able to test the sensitivity of our AuNP assay, for next year, we will be making sure that our system can detect up to 15.2 ng/mL of Her-2, and similar levels for Mucin-1 and Mammaglobin B(Zhang et al., 2020). Unfortunately, a set benchmark for the amount of Mucin-1 or Mammaglobin B that indicates breast cancer still does not exist, but we will adapt our assay to detect for both minuscule or large concentrations of the biomarkers to allow for its versatility.
Our planned tests for specificity and selection of aptamers allow for our test to only detect biomarkers specific to breast cancer. This minimizes the chances of false or inaccurate results as our aptamers bind specifically to our biomarkers and no other proteins. By selecting aptamers, we have not only developed a convenient way for detection but a method that is highly sensitive and specific.

Biomarker Collection Methods

When designing our testing device and our experiments, the first area of concern was our method of collection.
Clinical tests like mammograms or biopsies have proven to be extremely accurate and reliable for the detection of breast cancer. However, they are invasive and based on our extensive interviewing for human practices, many women are uncomfortable getting these tests and would prefer a more convenient method for detection. This is why we designed a testing kit that is convenient and portable to target our proposed audience, which was the main focus of our project.
Therefore, when we researched our biomarkers, we selected Mammaglobin B and Mucin 1. This is because Mammaglobin B is found in high levels in the lacrimal and salivary glands, while Mucin 1 is found in high levels in sweat gland cells. We hypothesized that since both biomarkers are present in sweat and tears, they are effective for a noninvasive method of breast cancer detection. Our final biomarker, HER-2, while found in blood, is an established biomarker with high efficiency, which is why we selected this as a comparison with Mucin-1 and Mammaglobin B.
Our idea for convenient detection was praised by Professor Kazunori Ikebukuro of Tokyo University of Agriculture and Science when we met with him to discuss our project idea, as he mentioned that some people are unwilling to undergo procedures such as the drawing of blood(even if it is painless) and that using alternatives such as detection through sweat and tears would be effective and life-changing. However, he cautioned us on methods of collection for sweat and tears, as biomarker concentrations can vary greatly. Based on his advice, we performed the following research for the collection of our biomarker in order to show how our project would be implemented in the market.

Collection of Mucin-1 Biomarker

Mucin-1 is found in high levels in sweat gland cells, and we hypothesize that through collecting sweat and analyzing for biomarker concentration, we will be able to detect levels of Mucin-1. Our research has shown that sweat can be collected through using pads or capillary tubing, and many commercial products exist on the market. Therefore, our solution is to collect sweat through a pad, filter the content, and using our Au-NP system to detect our biomarker.
Figure 2:Sweat Collection
Created with

Collection of Mammaglobin-B Biomarker

Mammoglobin-B is found in high levels in salivary and lacrimal gland cells. We hypothesize two methods to help collect samples for Mammaglobin-B.
First, we can collect salivary samples. With the COVID-19 pandemic, methods for salivary sample collection are very convenient and accessible. Additionally, saliva collection can be done at home without the use of specialized equipment or personnel, allowing for convenient at-home detection. However, when designing our collection system, we may need to consider methods to purify or monitor the quality of samples. Among the different methods for the collection of our biomarker, we hypothesize that this has the most potential due to the existing methods on the market and the prominence of this testing method. Therefore, we propose using a saliva collection technique for our physical testing device.
Figure 3:Saliva Collection
Created with

Another method would be to collect tears. However, this method has various limitations and considerations. First, depending on the type of tears, the content of biomarker may vary greatly and research on the content of Mammoglobin-B in lacrimal gland cells is still limited. In addition, while tear collection kits do exist on the market, they are not firmly established and there is a significant barrier with how to get individuals to cry to collect the tears. However, despite the limitations, we want to present this as a potential method for collecting our biomarker.
Figure 4:Tear Collection
Created with

Collection of Her-2 Biomarker

The concentration of HER-2 can be measured through a biopsy, but recent research suggests that there is enough concentration of HER-2 to be detected through blood.
Therefore, in order to collect HER-2, we recommend using an at-home blood testing kit, which widely exists on the market. However, at-home blood sample collection is prone to danger and can be potentially a safety hazard if users do not follow the procedures correctly. Nevertheless, blood sample collection is a prominent method for breast cancer detection due to the abundance of painless methods that exist. Combined with our Au-NP system, the sample collected from blood can be used to detect the amount of HER-2, and the presence of breast cancer can be determined based on the benchmark for HER-2. Therefore, we also propose using a blood collection technique for our physical testing device.
Figure 5:Blood Collection
Created with

At-Home Testing Kit

While our at-home detection device has not been designed, we envision that it will be used and implemented in the following manner.
First, with our collection process(blood or saliva), we will collect and purify our samples to be tested. These samples will be then added to our detection kit, which will consist of a tube that contains our aptamer-AuNP solutions that were synthesized prior to distribution.
Next, the user will add the sample to the Au-NP solution and add NaCl to induce the color change of the gold nanoparticles. By preparing the aptamer-AuNP solution beforehand, users will only have to do three steps—collect the sample through blood or saliva, add the sample to the prepared aptamer Au-NP solution, and add NaCl to induce the color change. These steps will take place in less than 10 minutes, allowing for quick and easy detection for users.
Figure 6: Example Sample Collection Data(Smith et al. 2016)
As you can see, there are different concentrations of biomarkers, and based on the different color hues, we can help derive the number of biomarkers for individuals using our test kit.
An alternative our team also developed was to extend our gold nanoparticle assay to a lateral flow strip. This would mean that we would construct a dipstick that employed the use of gold nanoparticles, similar in design to other products in the market such as HybriDetect.This models our initial solution to create a detection kit similar in design to a pregnancy kit. Upon addition of the biomarker sample, we can qualitatively and quantitatively analyze the color change to measure the concentration of biomarkers.
We will quantify the concentration of biomarkers by asking the user to take a picture of the color of their tube or strip. Through the use of our designed software, we will convert the color hue of our samples into the concentration of biomarker present and risk factors for breast cancer, allowing for users to get their results almost immediately.

Safety and Challenges

Since our test kit involves the collection of saliva or blood samples, proper disposal methods must be taken after the collection of samples to avoid contamination or risk of infection. Therefore, when designing our test kit, one challenge we face is proper methods to discard samples and we may need to discuss with our local government or community about appropriate measures.
Additionally, if our testing device employs blood collection, we will need to consider appropriate measures to help collect blood samples without harming or injuring the patient. We will do this by adopting or building on previously existing methods of at-home blood collection through using a capillary blood draw.
There are also various challenges that we would need to consider when implementing our testing device. First, we must determine a benchmark for the concentration of biomarkers needed to test positive for breast cancer. With the stigma existing against breast cancer patients being extremely high in Japan, having a false positive can create additional risk and anxiety for individuals. Therefore, our testing kit must be highly specific in measuring the concentration of biomarkers from the samples, and the threshold to test positive for breast cancer must meet local and global guidelines. Next, a second issue that may exist is cost. Gold nanoparticles are expensive to order. However, extending them to a lateral flow strip will greatly reduce the number of gold nanoparticles we need to use, marketing the detection kit at an accessible and affordable price.

Marketing and Distribution

Once we have verified that our test kit is able to be implemented in the market, the final concern is distribution and cost.
We plan on marketing our detection kit by selling it at local pharmacies or drug stores without prescription. We recognized that if we wanted to increase testing rates for breast cancer, our product had to be accessible to any person, just like any item one would buy at a drugstore. In addition, we plan on selling our product on online platforms such as Amazon to increase convenience and accessibility.
When we researched the possible production costs of our detection device, we looked into the already existing gold particle lateral flow strip, HybriDetect. For about 240 USD or 26,000 yen, individuals will be able to purchase 100 test strips. This means that each test strip is priced at around 2.40 USD or 300 yen, a price that is accessible and very reasonable. However, the production costs do not include the cost of reagents such as our aptamer(that needs to be conjugated) and biomarker collection devices. Therefore, factoring these into account, we estimate that our testing kit will be around 9 USD or 1000 yen.
However, we realize that selling our product in pharmacies or drug stores is not enough. Therefore, we propose that we collaborate with the government to distribute a testing kit to every household in Japan annually or monthly to allow for full-scale detection. This would be similar in design to the distribution of masks to every household by the Japanese government during the COVID-19 pandemic, with the costs of the mask similar in price to our testing kits. By distributing test kits to every household, we can ensure that women get tested for breast cancer on a regular basis, and seek help as early as possible to increase the chances of survival.


With our considerations for sample collection, test kit design, user input, and distribution, we believe that our project, ABrCaDaBra, could be implementable and marketable in the market as a method for breast cancer detection.
Presenting our magical solution, ABrCaDaBra!


  1. McKeague, M. (2017). Aptamers for DNA Damage and Repair. International Journal of Molecular Sciences, 18(10).
  2. Pan, R., Jiang, Y., Sun, L., Wang, R., Zhuang, K., Zhao, Y., Wang, H., Ali, M. A., Xu, H., & Man, C. (2018). Gold nanoparticle-based enhanced lateral flow immunoassay for detection of Cronobacter sakazakii in powdered infant formula. Journal of Dairy Science, 101(5), 3835–3843.
  3. Smith, J. E., Chávez, J. L., Hagen, J. A., & Kelley-Loughnane, N. (2016). Design and Development of Aptamer–Gold Nanoparticle Based Colorimetric Assays for In-the-field Applications. Journal of Visualized Experiments: JoVE, 112.
  4. Zhang, P., Xiao, J., Ruan, Y., Zhang, Z., & Zhang, X. (2020). Monitoring Value of Serum HER2 as a Predictive Biomarker in Patients with Metastatic Breast Cancer. Cancer Management and Research, 12, 4667–4675.