Overview

Our team, Resisense, is deeply committed to addressing the critical needs of our community and the world at large. To secure our project's success, we conducted extensive human practice interviews to understand the intricate details of detecting antibiotic resistance genes and the points of concern of implementing our kits in practice.

It is with sincerity that we say the success of our project is indebted to our human practice interviews. By investing time and effort into these conversations, the invaluable knowledge and broad perspectives we gathered enriched our project greatly, particularly in our wet lab and education events.

Interviews

Preliminary Project Design

Professor Karen Tam

Professor Tam is a research assistant professor in the chemistry department at the Chinese University of Hong Kong. She has extensive experience in functionalising biocompatible nanomaterials with theranostic agents for diagnosis and therapy, as well as in developing nano-devices for bio-analytics, computing, and other biological applications.

DNAzyme Feasibility

In the consultation with Prof Tam, we presented our initial design for discussion on its feasibility. Our original idea involved using DNA-cleaving DNAzyme that emits fluorescence upon binding to the targeted antibiotic-resistance gene. However, Prof Tam pointed out that the DNA-cleaving DNAzyme binding is not as specific as we had assumed, and its working temperature does not align with our goal of creating a point-of-care device.

Project Modification

As a result of this meeting, we have made significant changes to our project's mechanism. We realised our initial approach was overly optimistic regarding the biochemical reaction involved. Her opinion prompted us to modify the DNA sensing method by employing a specific enzyme and using G-quadruplex DNAzyme to indicate the presence of the targeted gene through a colour change in the later step.

Crux of Science Communication

Doctor Helen Ma

Doctor Ma is the science education project manager at the Office of University General Education at the Chinese University of Hong Kong and an RTHK television science programme host. She is an experienced science communicator, with a background in Streptococcus pneumoniae antibiotic resistance research in her post-graduate degree.

General Public's Understanding

Dr Ma expressed that while the general public is getting more educated on the issue of antibiotic resistance, not all people will take action to impede the spread of resistance. For instance, since medicine is often considered "harmful" and damaging to the body in the view of Hong Kong people, and antibiotics are only administered when the symptoms are severe, people often choose not to complete their antibiotic prescription. The concept of antibiotic resistance is vague and distant to the general public, giving them low motivation to take the right action.

Effective Science Communication

We also consulted her on the best practices to effective science communication. Dr Ma suggested that when describing abstract ideas to people, especially the older generation, we need to emphasise how it will directly affect them instead of how it works. For instance, instead of explaining how the leak of antibiotics leads to the development of resistance, we should directly tell them that their medicine will not work next time if they don’t complete their prescription and that doctors will eventually run out of effective medication. It is more effective to convince them by illustrating how they will be directly affected instead of lecturing them on the mechanisms and causal relationships.

Project Modification

Her precious opinion shed light on our education work. She pointed out how the public misunderstands antibiotics, leading to their misuse and abuse. We thus aimed our workshops towards offering correct information on antibiotics and introducing them to raise public awareness. Her opinion was taken into great consideration when designing our CUHK art fair game.

Probe Vector Selection

Professor Margaret Ip

Professor Ip is the Chairperson of the Department of Microbiology at the Chinese University of Hong Kong. She is an expert in Clinical Microbiology and Infection and is a renowned member of various antibiotics-related committees. 

Market Analysis

In the interview with Prof Margaret Ip, we gained a comprehensive understanding of the current situation of antibiotic resistance in Hong Kong and the corresponding detection methods. Prof Ip explained that the most common way to detect antibiotic resistance genes is through DNA sequencing with protein analysis. However, this method is expensive, especially in developing countries. It requires a lab expert and a large machine to analyse.

Probe Characteristics

Prof Ip suggested that DNA is more stable and sensitive than detecting proteins or toxins produced by bacteria alone. She recommended that we detect multiple genes in one test for the sake of sensitivity and accuracy due to the occurrence of multiple genes in bacteria. We plan to detect related DNA to infer antibiotic resistance in targeted bacteria. Proteins and toxins will be our alternative if the DNA sensing approach does not work in our web lab. As per Prof Ip's emphasis, our point-of-care device should be cheaper and faster than DNA sequencing techniques currently used. 

Project Modification

Due to the relative stability of DNA molecules compared to protein, we confirmed that we have to detect the desired antibiotic-resistant gene instead of protein and toxin. Before the interview, our team reviewed various DNA sensing methods, including Rolling Circle Amplification (RCA). This interview led us to focus on DNA molecules to increase sensitivity and effectiveness. Following this interview, we confirmed we would employ RCA as our primary detection method. 

Point-of-care Kit Design

Professor Stephen Tsui

Professor Tsui is a renowned expert in the field of Biomedical Sciences and the director of the Hong Kong Bioinformatics Centre and the Centre for Microbial Genomics and Proteomics at the Chinese University of Hong Kong.

Device Reliability

During our conversation, Prof Tsui provided valuable insights on our project design and future goals, highlighting some key points we had overlooked, such as the issue of specificity and accuracy, including false positive/negative problems and purification issues. He suggested that impurities could lead to significant problems in the accuracy and sensitivity of our RCA step. To address these issues, Prof Tsui recommended some purification steps and implementing controls in our test to improve the reliability of the results.

Targeted Group

Prof Tsui also shared his precious opinions on our targeted group, pointing out that the most popular and mature approaches to detect antibiotic-resistant genes nowadays in developed countries include MIC, DNA sequencing, real-time PCR, and single-point mutation enzymes. However, these techniques are uncommon in developing countries, so we could be more competitive if we target them.

Results Display and Verification

Additionally, Prof Tsui suggested that our project design tends to be qualitative rather than quantitative, like MIC. He explained that a qualitative approach might not be ideal for clinical use, as doctors may need help determining the result and taking appropriate actions based on it. Therefore, he recommended that we decide how to show a colourimetric result in the last step, such as coupling a digital technology with the POC device to detect the colour change or a COVID-RAT-like approach of showing two lines. (See Our Solution)

Project Modification

Prof Tsui's experience provided imperative insight into the results display and our proposed hardware design for point-of-care kits. We designed a colourimetric result display in the final step using G-quadruplex DNAzyme, targeting a similar approach to the COVID-19 rapid test kit. We decided to provide a qualitative result, indicating only "yes" or "no" for a more straightforward interpretation of the result. Suggestions from Prof Tsui also enabled our team to arrange a purification step in the test kit to prevent the residue from the previous steps from affecting the next step.

Frontliner's Perspective

Doctor Pierre Chan

Doctor Pierre Chan is a medical practitioner and a former legislative counsellor seated as the representative for the medical functional constituency in Hong Kong.

Hospital Procedure

Dr Chan explained that in severe cases, doctors usually use empirical antibiotics based on international guidelines to save lives instead of finding the most appropriate drugs. However, in non-severe cases, doctors check the type of bacteria involved in the infection. He also mentioned that several samples such as urine, blood, and stool are collected from patients, but only 2-3 samples usually show positive results. It is crucial to consider the type of samples collected for analysis. Furthermore, hospitals usually do not recheck the results due to time and manpower constraints, and a full analysis takes 1-2 days.

Public Awareness

Dr Chan believes that the problem of antibiotic resistance is more prevalent in the community than in hospitals. He cited the misuse of antibiotics in the agricultural industry as an example of how the abuse of these drugs is not regulated. Although public awareness about antibiotics is not low in Hong Kong, Dr Chan pointed out that some people buy restricted drugs, including antibiotics, without a doctor's prescription. Our discussion highlighted the need for educational events to address this issue. It is essential to consider different approaches to education to ensure that the public is well-informed about the importance of responsible use of antibiotics.

Project Modification

Our discussion with Dr Chan shed light on the use of antibiotics in hospitals and the public awareness about antibiotics. We will focus our point-of-care kit for screening speed. We will also consider his insights while developing our educational events to ensure that the public is well-informed about the importance of responsible use of antibiotics.

Biotechnology Education in Local Secondary Schools

Dr Victor Lau

Doctor Victor Lau is the assistant dean of the Department of Curriculum and Instruction at the Chinese University of Hong Kong and a consultant for the science education curriculum in Hong Kong.

Biotech Development in Local Secondary Schools

As local students, we have experienced Hong Kong’s science education and its flaws. Biotechnology is currently an elective topic in the biology curriculum, covering fundamental theories like PCR and gel electrophoresis. However, as mentioned by Dr Lau, it is not cost-efficient for a secondary school to invest in a complete set of biotechnology lab equipment solely for teaching a few chapters. Targeting this problem, Hong Kong Amgen Biotech Experience (ABEHK), an organisation co-directed by Dr Lau, is supporting local schools by renting out equipment like thermocyclers and pipettes to schools and sending student ambassadors to provide hands-on experience to secondary school students, similar to typical iGEM secondary school experience workshops.

Education on Antibiotic Resistance

Dr Lau suggested focusing more on antibiotic resistance in our education workshops. He pointed out many misconceptions on this matter, not only among students but also among the general public. The most common one is that resistant bacteria only come from the use of antibiotics. He also highlighted a trend of medical malpractice where local clinics are not giving the full antibiotic dose to cut costs, leading to resistant bacteria development. Although the government has distributed posters and leaflets on this topic, the message is not well-absorbed. Dr Lau suggested using simple interactive models assembled with simple stationery like paper clips and magnets to explain relevant concepts to the public in an interactive way over a verbal-only approach.

Project Modification

We pointed out that one of the most common issues secondary school students face is that they need to troubleshoot experiments. Workshops like those provided by ABE allow students to understand better the theory and, hence, the errors that led to their failures. After learning about ABE and its mission, we planned to build upon their workshops and design one of our own, specifically with material to teach students how to troubleshoot.

International Regulations and Opportunities

Dr Edmond Ma

Doctor Edmond Ma is the AMR consultant for the Infection Control Branch at the Centre for Health Protection of Hong Kong.

Global AMR Monitoring

During our interview, Dr Ma introduced us to the ever-updating Bacterial Priority Pathogen List (BPPL) published by the World Health Organisation (WHO). Every year, the WHO refines the list by highlighting bacteria that can resist antibiotics and categorises them into three groups according to their threat level. The list aims to notify doctors of the appropriate treatment that should be taken for cure while also projecting a trend of emerging antibiotic resistance for researchers for reference when developing new ways to combat the bacteria. Meanwhile, the WHO also provides guidelines for the use of antibiotics in the form of Antimicrobial Stewardship Programmes (ASPs). This serves as an education for doctors and health professionals not to abuse antibiotics and antimicrobial medicines, as the overuse of such drugs will accelerate the development of antibiotic resistance in bacteria.

AMR Outside Hospitals

The abuse of antibiotics can also happen outside of the typical health workplaces. As Dr Ma mentioned, wastes containing antibiotics might be released from various sources, such as farms that use antibiotics as growth promoters for crops and livestock. When these contaminants enter natural rivers or seep into nearby soils, the local bacteria will be exposed to antibiotics unintentionally. While antibiotics might kill some bacteria, such action will only facilitate the emergence of antibiotic resistance as the bacteria attempt to survive.

Opportunities for Our Project

On the design of a point-of-care antibiotic resistance detection kit, Dr Ma agreed that it could help safeguard doctors and patients in a remote or private clinic setting, especially when the patient is in the early stages of infection where only broad symptoms are shown without a clear indication of the type of infectious bacteria. On the other hand, Dr Ma questioned whether a false positive would be detected when the patient is a mere carrier of the resistant bacteria, as a concrete connection between the detected gene and the bacteria causing the infection has yet to be established. It is suggested that an analysis of the rate of carriers of a particular bacteria in the general public should be conducted to estimate the kit's accuracy.

User Experience

Other than that, Dr Ma commented on a few aspects that should be considered when designing a testing kit, one of which is the user experience. A well-designed kit should minimise the suffering of patients during use, including relatively low or no aftermath, such as nausea or aching of body parts after use and pain during sample extraction. Regarding patient samples, Dr Ma also noted that the sample type should be specific to the illness, as a disease-causing bacteria in one body part might be relatively harmless in other locations. For example, a bacteria might induce coughing when it is present in the lungs while having no effect when it enters the intestines, so the sample taken should be products from the respiratory system, such as phlegm, rather than excrements from the digestive system as a presence of the bacteria in such samples can not prove that that causes the coughing.

Project Modification

After the interview with Dr Ma, we understood that multiple countries are working on curbing the dissemination of antibiotic resistance, and many low- and middle-income countries are facing the consequences. Dr Ma pointed out that many doctors use so-called big-gun antibiotics for suspected bacterial infections before knowing the MIC result. Besides, he mentioned that many developing countries cannot afford the exorbitantly expensive PCR machine for AMR checking. Therefore, we can have a place in the market to provide cheap and fast testing methods to fill in the gap. This interview gave us a new direction for selecting our target market and our aimed application.