Team:YiYe-China/Implementation
Implementation:
When it comes to CRC detection methods, people always first thought of colonoscopy, CT scan, etc. But, such testings not only have severe side effects and discomfort, they are also expensive and not convenient. Therefore, we want our project to be as close to commercially viable as possible so that everyone could have access to it. We constantly referred back to our human pracitce research, which influenced all the way from construct design to the marketing plan, to guide the implementaiton of our project.
Who are our proposed end users?
CRC can be divided into different stages (eg. Stage I, Stage, II, Stage, etc.), based on the severity of the patient's situation. But fortunately, our project could be applied to all stages of CRC. Indeed, the target DNAs used in our method have an accuracy rate of 76%--89% among all stages of CRC. Therefore, our target users are every person potentially having cancer polyps.
But since certain processes require a specifc training, for instance, the use of pippetes, we still recommend professional people, like laboratoary doctors, get to do the procedure. Though extracting DNA from stool sample sounds easy, we still have to follow the standardized rule to ensure the accuracy, For instance, Urinalysis, one of the simplest screening test, still requires professonal people. Hence, the end users should be someone with a bsaic laboratoay experience.
How do we envision others using our project?
We envision our whole process being convenient for patients. We first refine the DNA from the stool sample collected from patients. Then, we add bisulfite to the samples for conversion. Bisulfite can turn the c-base to the u-base. However, when the DNA is methylated, the c-base will retain despiting adding bisulfite. So we could tell the samples’ statuses by seeing if the base is transformed. Next, through PCR amplification, we got our Target DNA and trigger. To ease the process, we could even use RPA amplication as a subsitution. Finally, we add our PCR product as well as our p1 plasmid toehold into the cell-free system and incubate for 3--4 hours. Thanks to the toehold switch, which has a a Red Fluorescent Protein mCherry binding at its tail, when the trigger DNA is detected, in our case, the methylated DNA, we are able to tell the methylation status of our sample through naked eyes.
The biggest highlight of our project is its being non-invasive and cost-effective. As many people complain about the discomfort the feel during the colonoscopy tests, our project can significantly lessen the discomfort, and there will be no side effect.
Also, our product requires the basic equipments so that clinics or community hospitals in the undevloped places also have access to the apparatuses for detection. In other words, patient no longer need to travel all the way to the hospitals in the city center. This could reduce the inconveniency of testing in big hospitals and also decrease the time waiting for screening.
In addition, people could even get their colon and rectum checked when doing physical exam as the process gets simplified to be implemented, thus allowing more people to get their cancer detected earlier.
How would we implement our project in the real world?
When we interviewed Dr. Li, an expert in molecular genetics, he suggested us using SDC2 as a target DNA in addition to TFPI2 and conducting a combined genetic test to see which gene would be the most effective in detecting. And if they both showed similar results, our project could be more accurate. So, for our future implementation, based on what Dr. Li suggested, we plan to do an experiment with SDC2 to increase our accuracy.
To apply our project to the public, we would also collaborate with BIT and combine our project with their equipemnt. BIT was going to create a mini app on Wechat and use bluetooth to detect fluoresence proteins and multilple different channels' movements. Then, the data would send back to bluetooth to show if there is any polyps detected. Though our teams used different fluorescent protein, we may either change our protein or add a filter to change the fluorescent color so that BIT's equipment could also be used to detect our fluorescence values.
The safety aspects we would need to consider
When we did our experiment, we had to follow the safety protocals in laboratory accept safety training before experiment. Since we were working with E. coli, we were required to wear lab coats, gloves and masks before entering the laboratory, and when we were done with E.coli., we would sterilize the culture by reasonable ways, like high temperature or high pressure and dispose all the waste properly. We were also equipped with quick first kits, such as eye washers to protect ourselves if anything unexpected happened.
But when applying to the real world, our product is relatively save. The strains of E.coli we chose are classified as low risk. There was no acid release or any significant danger that we need to pay attention to.
Therefore, we could ensure that when our product is applied to the public, every person, including those with high risk facotrs, should be able to use it without any concern.
The challenges we would need to consider
1. The first challenge is to increase the sensitivity and the accuracy of our project. This is what people value the most when it comes to diagnostic tests, and we also strive to increase the two values. However, due to the conditional restrictions, we couldn't do clinical trials. But we will add another target DNA, SDC2, in addition to the one we used (TFPI2) to improve the accuracy. When we compared the results we get from both the two DNAs, if they show similar results, it means that our product's sentivity could reach up to 98% and have a 95% accuracy.
2. The second one is to raise the awareness of our product. When we sent out survey about our projet, though the majority are supportive of our project, there was still a group of people who remain skeptical. Therefore, to increase the credibility of our project is another challenges. We remembered that when Dr. Sheng (the doctor we interview) first tried to use a steel needle to eliminate a liver cancer. Compared to regular methods, such as chemotherapy or radiation therapy, the technique of microwave thermal ablation at that time was rare, and many people opposed taking such a big risk. However, the results turned out to be well, and that changed people's mind. Our project may face the same challenge as Dr. Sheng faced. There may be people who find it difficult to accept such a novel method. Though our project only plays a role of aided detection, people may still refuse to use it. To resolve the issue, we could make some posters and host some workshops to let people understand more about our project. By emphasizing that our project is non-invasive and convenient to use, we hope that more people would be more flexible with this new method.
3. The third challenge is the our product's competency when compared to FOBT. When people think of non-invasive detection test, fecal occult blood test (FOBT) will first pop out into their minds. However, FOBT has a low accuracy rate: their sensitivities are relatively low in detecting stage I CRC (53%) and advanced adenomas (≥1.0 cm) (27%), with a specifi- city of 94.6% The test could show a negative test result when cancer is present (false-negative result) if your cancer or polyps don't bleed. On the other hand, the test could show a positive result when you have no cancer (false-positive result) if you have bleeding from other sources, such as a stomach ulcer.
Therefore, we have to eliminate people's stereotype of such detection methods as they may think of our project less sensitive as well. To elimiate people's misunderstanding, we should let more people know the differentce between out project and FOBT. In addition, we should stress out the accuracy rate of our project.
Reference:
[1]“Fecal Occult Blood Test.” Mayo Clinic, Mayo Foundation for Medical Education and Research, 12 May 2020, www.mayoclinic.org/tests-procedures/fecal-occult-blood-test/about/pac-20394112. [2]Morikawa T, Kato J, Yamaji Y, Wada R, Mitsushima T, Shiratoti Y. A comparison of the immunochemical fecal occult blood test and total colonoscopy in the asymptomatic population. Gastroenterol. 2005;129(2):422–8. [3] https://www.peoplematters.in/article/employee-relations/the-high-performance-organizational-framework-and-how-to-implement-it-23563
Implementation
Our Proposed Implementation
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End users
Who are our proposed end users?
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Envision
How do we envision others using our project？
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Implementation
How would we implement our project in the real world?
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Safety
The safety aspects we would need to consider
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challenges
The challenges we would need to consider
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Reference
The papers we referred