Korea may be famous for internet startups and K-Pops, but not for synthetic biology. Synthetic biology has a huge potential for improving the welfare of humanity, but being such a young discipline, at the same time has many issues yet to be resolved, including ethical, legal concerns and safety issues. In US or EU, thus, it is not uncommon to see close cooperation between the government and private enterprises to create an ideal research environment and efficient legal and regulatory framework.

No such environment exists in Korea, at least up to the level of efficiency found in US or EU. We have concluded that the policy makers and important players (such as colleges and research institutes) must be better informed of the new field so as to provide an environment that is favorable for the advancement of synthetic biology while safe enough to prevent hazardous events. We have also identified that it is necessary to promote fundamental concepts of synthetic biology to younger generations, including high school students and younger. Keeping these social environment of Korea in mind, Elan Vital Korea has established 5 goals for our human practice activities:
- first, enhancing public awareness of the synthetic biology;
- second, promoting the importance of the field to policy makers, research institutes and journalists;
- third, promoting the public awareness of the importance of the early detection of contagious pathogens;
- fourth, putting safety first for our projects;
- and finally, close cooperation and co-work with other iGEMers in Korea.

First, we continuously and strenuously strived to enhance the public awareness of the synthetic biology, especially among younger generations. We have participated in Korea Youth Expo and operated a booth (from May 22 to May 24), and worked hard to communicate with participants in 2015 Science Festival (from July 27 to August 2). We offered basic education sessions on DNA and synthetic biology, and conducted surveys to better understand the level of general awareness of the synthetic biology and iGEM. Based on the information and data we gathered from them, we have developed the long-term education plan for primary and secondary school students. We have reached the National Science Teachers’ Association of Korea, with more than 2,400 science teachers as its members, and established a communication and cooperation network. We have provided some materials for use for education including blogs, Youtube videos, lecture videos, booklets, brochures and simple DNA extraction kits, etc.

Second, we have learned that policy makers and education and research bodies in Korea are not sufficiently informed of the recent developments of the synthetic biology, which resulted in lack of proper safety guidelines and articulation of legal, ethical and environmental implication-related discussions and awareness. In an effort to change the condition, we have drafted the report of our findings and suggestions, and sent our concerns and recommendations to politicians (including members of the National Assembly of Korea) in charge of science and technology-related policymaking. In the process of researching the current policy environments and developing recommendations, we have contacted universities, research institutions and small- and medium-size enterprises in order to better understand their concerns. We have also contacted journalists, who function as major contact point to the general public. We have distributed summaries of the synthetic biology and iGEM. In the process, we have persuaded some of them to be our sponsors.

Third, we have developed the public awareness strategy for our project, “Early Detection of Antibiotic Resistant Bacteria Using Quorum Sensing”. Incidentally, Korea has been hit hard by MERS virus as late as June this year, which greatly heightened the public awareness – and fear – of the contagious pathogens, as well as the need to learn more of the cause and cure for those dreadful diseases. We began our public awareness campaign by researching and studying the current methods of detecting and tackling the antibiotic-resistant bacteria from Korean CDC, hospital guidelines and related research papers. We have discovered that, compared to the strict and detailed action plan in place by the US CDC, the Korean CDC has not – yet – established networks of hospitals and nursing homes that need to be closely monitored and promptly intervened as need arises. Based on this finding, we have drafted letters and papers and sent them to Korean CDC and safety committees of hospitals. We plan to continue our efforts on this area.

Fourth, we have put the strongest emphasis on the safe implementation of our project. Our project belongs to Risk Group 1, but we have observed the lab safety rules and guidelines very tightly, and promoted the importance of observing the lab safety based on deeper understanding of the biosafety guidelines. We have developed the Youtube video explaining the biosafety levels, and encouraged people in lab environment to watch it. Finally, in an effort to promote the spirit of cooperation, sharing and mutual contribution of iGEM, we have reached out to other teams joining the iGEM Jamboree, and hosted regional meet-up. Even before and after the regional meet-up, we held discussions and mentorship with Korea University team on lab works and iGEM experiences. We also met with HAFS team and provided assistance on parts submission and using BioBricks and other materials included in the Kit. We have shared with them our failure and misunderstanding we had experienced last year which, we hope, would be entertaining if not informative.

The spread of antibiotic-resistant bacteria is a global health problem that affects nations across borders and boundaries and rapidly spreads throughout the world. These nightmare bacteria pose catastrophic threat to people everywhere in the world.

Our project is Early Detection of Antibiotic Resistant Bacteria using Quorum Sensing. It is closed related to the surveillance and containment of the pathogens. With the understanding, we have conducted case study of Korea and the US on the current status of antibiotic resistant bacteria infection and the countermeasures. We have gathered data on the spread of antibiotic-resistant bacteria and countermeasures taken as well as contemplated in Korea and the US. For this, we have relied on various materials and documentations, and some of them are translated from Korean.

Available data are insufficient to estimate the wider societal impact and economic implications when effective treatment for an infection is completely lost as a result of resistance to all available drugs. The overall health and economic burden resulting from acquired antibiotic resistant bacteria cannot be fully assessed with the presently available data; new methodologies are needed to more precisely assess the total impact of resistance, to better inform health policies and to prioritize the deployment of resources. However, even admitting the lack of reliable information on the financials, the overall cost is highly burden to all nations, even further to the less developed countries. For example, the yearly cost to the US health system alone has been estimated at US $21 to $34 billion dollars, accompanied by more than 8 million additional days in hospital. Because antibiotic resistant bacteria has effects far beyond the health sector, it was projected, nearly 10 years ago, to cause a fall in real gross domestic product (GDP) of 0.4% to 1.6%, which translates into many billions of today’s dollars globally.

In Korea, Korean CDC collected antibiotic-resistance infection cases from hospitals, which shows that 41,883 patients (about 7% of the total hospitalized patients) were infected by antibiotic-resistant bacteria during the first half-year of 2014 (from January 1 to June 30, 2014).

(Source: Korea Center for Disease Control and Prevention, http://www.cdc.go.kr/search/sEngine.jsp?kwd=MRSA%EA%B0 %90%EC%97%BC)

KARMS released annual report in 2014 containing antibiotic resistance infection data gathered from hospitals as well as long term care facilities and small & medium sized clinics. Publishing such data is a clear evidence that the Korean government is keenly aware of the hazards of the antibiotic resistant bacteria and also shows its commitment to deal with the matter systematically. The government adopted a law on the Prevention and Containment of Infectious Diseases in Dec. 2010.

In accordance with the executive order of the law, antibiotic resistance infections including VRSA, VRE, MRSA, MRPA, MRAB, CRE were designated as “infections to be contained”. The annual reports deals with S. aureus, Enterococcus spp., S. pneumoniae, E. coli , K. pneumoniae , E. cloacae, P. aeruginosa , A. baumannii Following are the results: (Data from General Hospital (more than 300 beds))

Medium Sized Hospital
Table * Antimicrobial resistance rates (%) of S. aureus isolated from hospitals

* Including cefoxitin, † Not tested, ‡ Trimethoprim-Sulfamethoxazole, , ∥ Quinupristin-Dalfopristin

As shown on the above data, among gram positive bacteria, antibiotic resistance of S. pneumoniae against penicillin G increased substantially in 2010 and reduced by 5.6% in 2012. On the other hand, antibiotic resistance of gram-negative bacteria has not much changed except A. baumannii that recorded 71.1% of resistance rate in 2010 and moderately decreased to 69.5% in 2012.

Vancomycin resistance rate of VRE E. faecalis is much lower than that of E. faecium, which is less than 1.0%. Vancomycin resistance rate of E. faecium at the long-term care facilities is rapidly increasing from 20.5% in 2007, 41.7%, 2009, and 55.6% in 2012 respectively. Vancomycin resistance rate at the small clinics is 23.9% in 2012, increased more than twice from 2010, higher than that of hospitals (22.0%) Vancomycin resistance rate of all Enterococcus spp. (including E. faecalis and E. faecium) shows similar level at hospitals and clinics in 2011. However the ratio at the long-term care facility is in gradual increase, registering 23.8%.

Antibiotic resistance rates of all medical care providing institutions have shown steady decrease from 2007. In 2007, among 5373 Patients of government operating health centers 16.1% have shown antibiotic resistance and 5.5 % multi-drug resistance. Afterward, the ratios have been slightly decreased each year. Meanwhile antibiotic resistance of private health care institutions is much higher: 29.1% in 2007, 19.8% in 2008, 23.6% in 2009, 21.4% in 2010, and 18.4% in 2011 respectively.

MRSA Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most severe antibiotic resistant pathogens causing hospital infection and other diseases such as purulent infection, bacteremia. The isolation ratio of MRSA is gradually increased up to 80% in the hospital, which makes a limitation for treatment of antibiotics because the isolated MRSA show resistance to methicillin as well as other antibiotics. To deal with the spread of MRSA infection, medical community as well as the government have paid acute attention on the surveillance of the infection.

Common used method for the detection of MRSA in the hospitals is Susceptibility Test. Another detection method is MecA gene by PCR, which is not commonly used in the hospitals because of the cost consideration. However mecA is known to be more accurate than Susceptibility Testing.

1. Taking Sample

1)Take secretion from infected skin with sterilized swap.
2)Taking phlegm from a potential patient or for those who cannot cough by themselves because of respirator, taking samples by cleansing of respiratory organ or bronchoscopy.
3) Taking urine sample from a potential patent or poley-catheter.
4) Taking blood sample.


2. Conduct susceptibility test

It usually takes 48 hours for a patient to get the result. In case
that the patient asks faster detection, other detection method
such as PCR, DNA sequencing are recommended. The
alternative test can produce the result within a few hours.

MIC (Minimum Inhibitory Concentration) is most commonly used susceptibility test. It is used for the detection of all pathogens. However, pathogens such as Streptococcus, Haemophilus, Neisseria that are hard to be cultivated are not detected by MIC. Mcroscan system that is using automated microdilution and E-test are used for MIC.

Paper Disk Method For the test, spread plate culture and pour plate culture are mostly used. Depending on the type of LB agar plate and testing antibiotics, many manufactures have been offered a variety of testing kit. This method should be done by manual procedure from the beginning to the end. Therefore standardized reliable protocols of the testing method should be developed and well observed.

The antibiotic to be tested is added to agar, which is then placed in dilution plates and diluted with varying levels of water. After this, the pathogen to be tested is added to each plate, plus a control plate that does not receive any antibiotics. The dilution plates are then incubated at a temperature of 37 degrees C. The plates are then incubated for sixteen to eighteen hours, although incubation time may be less for bacteria populations that divide quickly. After incubation, the plates are examined to determine if bacterial expansion has occurred. The lowest concentration of antibiotics that stopped the spread of the bacteria is considered to be the minimum inhibitory concentration of that bacteria

Agar dilution is considered to be the standard of susceptibility testing, or the most accurate way to measure the resistance of bacteria to antibiotics. The results of agar dilution are easily reproduced and they can be monitored at a much cheaper cost than what is required of other dilution methods. Additionally, up to thirty pathogen samples (plus two controls) can be tested at 0o0nce, so agar dilution is useful for batch tests.

Each dilution plate in agar testing has to be manually infected with the pathogen to be tested, so agar dilution testing is both labor intensive and expensive. And agar dilution cannot be used to test more than one pathogen at a time.

The tube dilution test is the standard method for determining levels of resistance to an antibiotic. Serial dilutions of the antibiotic are made in a liquid medium which is inoculated with a standardized number of organisms and incubated for a prescribed time. The lowest concentration (highest dilution) of antibiotic preventing appearance of turbidity is considered to be the minimal inhibitory concentration (MIC). At this dilution the antibiotic is bacteriostatic.

Varying concentrations of the antibiotics and the bacteria to be tested are then added to the plate. The plate is then placed into a non-CO2 incubator and heated at thirty five degrees C for sixteen to twenty hours. Following the allotted time, the plate is removed and checked for bacterial growth. If the broth became cloudy or a layer of cells formed at the bottom, then bacterial growth has occurred.

The broth dilution method can be used to test the susceptibility of bacteria to multiple antibiotics at once. Broth dilution is also highly accurate. The Other advantages include the commercial availability of plates, the ease of testing and storing the plates, and the ability for the results of some tests to be read by machines. However, the broth dilution method is only effective for testing Bacteroides fragilis. It could potentially be used with other bacteria populations, but the results would have to be correlated with agar dilution tests to be considered reliable. Although the tube dilution test is fairly precise, the test is laborious because serial dilutions of the antibiotic must be made and only one isolate can be tested in each series of dilutions.

Beta-Lactamase Test is a means of detecting the enzyme beta-lactamase, which confers penicillin resistance to various bacterial organisms by cleaving the beta-lactam ring of penicillin and cephalosporin antibiotics. A wide variety of bacteria produce this enzyme, including both gram-positive and gram-negative organisms. This acidimetric method is recommended for use in testing beta-lactamase production by Neisseria gonorrhoeae, Haemophilus species and Staphylococcus species.

Etest, (previously known as Epsilometer test) manufactured by bioMérieux, is a manual in vitro diagnostic device used by laboratories to determine the MIC and whether or not a specific strain of bacterium or fungus is susceptible to the action of a specific antimicrobial. This type of test is most commonly used in healthcare settings to help guiding physicians in treatment of patients by indicating what concentration of antimicrobial successfully would treat the infection.
MIC test has been used as standard test method of the detection of antibiotic resistance in hospitals. It has weaknesses such frequent error caused by agar LB, disk, tested pathogens, reference strain, and cultivation condition, and lab technician.

Another shortcoming of MIC test is the time to get the result. In terms of accuracy and required time, other advanced tests such as mecA PCR Method is recommended by some hospitals. But still majority of detection is being by MIC.

Actual cost needed for the detection varies hospitals from hospitals. However average cost of susceptibility test is 55000KW ($50) to 90000KW ($85). For other test, the testing cost is more than 250000KW ($ 200). Testing expense is not covered by mandatory government insurance plan. Thus, if the patient does not have private insurance policy, she should pay by herself.

The CDC reported that each year in the United States only, at least 2 million people acquire serious infections with bacteria that are resistant to one or more antibiotics designed to treat the very infection. At least 23,000 people die each year as a direct result of these antibiotic-resistant infections. Many more die from other medical conditions that were further complicated by an antibiotic-resistant infection. (Source: Antibiotic Resistant Threats in the United States 2013, Center for Disease Control and Prevention)

In order to cope with the threats, CDC is adopted five major tasks in order to support initiatives of the White House’s National Strategy to Combat Antibiotic Resistant Bacteria (https://www.whitehouse.gov/sites/default/files/docs/carb_national_strategy.pdf)and the President’s Executive Order(https://www.whitehouse.gov/the-press-office/2014/09/18/executive-order-combating-antibiotic-resistant-bacteria)

 Slow the Development of Resistant Bacteria and Prevent the Spread of Resistant Infections
 Strengthen National One-Health Surveillance Efforts to Combat Resistance
 Advance Development and Use of Rapid and Innovative Diagnostic Tests for Identification and Characterization of Resistant Bacteria
 Accelerate Basic and Applied Research and Development for New Antibiotics, Other Therapeutics, and Vaccines
 Improve International Collaboration and Capacities for Antibiotic Resistance Prevention, Surveillance, Control, and Antibiotic Research and Development

In order to fully implement the National Strategy for Combating Antibiotic-Resistant Bacteria, the FY16 AR Solution Initiative was adopted. This implementation will include: comprehensive tracking and detection of antibiotic-resistant bacteria, faster outbreak response, insights for research innovation, better patient care, improved prescribing, increased susceptibility testing, nationwide implementation of CDC’s Core Elements of Hospital Antibiotic Stewardship (http://www.cdc.gov/getsmart/healthcare/implementation/core-elements.html), and global partnerships for prevention and detection.

The FY16 budget also supports a $14 million increase for the National Healthcare Safety Network (NHSN)—the nation’s leading system to track healthcare-associated infections, including antibiotic resistance and antibiotic use—as a companion to CDC’s FY16 Antibiotic Resistance Solutions Initiative, supporting multiple goals under the National Strategy, including new activities to better understand and monitor sepsis, leading to enhanced prevention to save lives.

One of the elements of the initiative is advance development and use of rapid and innovative diagnostic tests for identification and characterization of resistant bacteria. FY 16 AR Solution initiative also includes comprehensive tracking and detection of antibiotic-resistant bacteria.

According to the National Action Plan to Combat Antibiotic-Resistant Bacteria released on 27, March, 2015, Improved detection and control of antibiotic resistance in human and animal pathogens will be achieved through a “One-Health” approach to disease surveillance that integrates data from multiple monitoring networks. To achieve the goal by 2020 the government has adopted action plan in order to generate significant outcomes.

Creation of a regional public health network—the Detect Network of AR Regional Laboratories—for resistance testing, a specimen repository for resistant bacterial strains, and a National Sequence Database of Resistant Pathogens.

Routine reporting of antibiotic use and resistance data to National Health Safety Network (NHSN) by 95% of Medicare-eligible hospitals, as well as by Department of Defense and Veterans Affairs healthcare facilities.

Routine testing of zoonotic and animal pathogens for antibiotic susceptibility at ten to twenty National Animal Health Laboratory Network (NAHLN) and Veterinary Laboratory Investigation and Response Network (Vet-LIRN) member laboratories, using standardized testing methods and data sharing practices.

Also important is advance development and use of rapid and innovative diagnostic tests for identification and characterization of resistant bacteria. In order to facilitate the achievement of the task, the government will take the lead with the support of funding agencies such as the National Institutes of Health. When it is achieved researchers are taking advantage of new technologies to develop rapid “point-of-need” diagnostic tests that can be used during a healthcare visit to distinguish between viral and bacterial infections and identify bacterial drug susceptibilities — an innovation that could significantly reduce unnecessary antibiotic use. The availability of new rapid diagnostic tests, combined with ongoing use of culture-based assays to identify new resistance mechanisms, will advance the detection and control of resistant bacteria.

By 2020, significant outcomes in this area will include:

Development and dissemination of authorized point-of-need diagnostic tests that rapidly distinguish between bacterial and viral infections.

Validation of diagnostic tests that rapidly determine the antibiotic resistance profiles of bacteria of public health concern.

As proved by these efforts, it is critical to detect antibiotic resistant bacteria as early as possible and eventually to develop “point-of-need” diagnostic test. Our project is to contribute to the early detection of antibiotic resistant bacteria by using quorum sensing. We have succeeded to create plasmids to detect antibiotic resistant bacteria though we are aware that it is rudimentary development of such kind but we believe it is used as a proof of concept and with advanced research efforts, it can be further developed to an extent that it can be used at the health care institutions and nursing homes. Our hope is to develop a simple kit just like diabetic test kit, so that anyone who needs detection of antibiotic resistance can use it.

Current detection methods require complicated procedure. As described in our background of project.(https://2015.igem.org/Team:elan_vital_korea/background#) Consequently, Antibiotic-resistant infections add considerable costs to the US healthcare system that can be avoided by devising ways to diagnose and treat those infections. Generally, patients suffering from antibiotic-resistant infections require prolonged, more expensive treatments, stay at hospitals longer, demand additional doctor visits, attention and care, and result in greater disability and death than those who were infected but can be easily treated using antibiotics. The total economic cost of antibiotic-resistance to the US economy is difficult to calculate, but is estimated to be as high as $20 billion of direct healthcare costs, plus additional burden to society for lost productivity of about $35 billion a year. (source: CDC). Unfortunately, information on unit cost for detecting antibiotic resistant bacteria is not available. We have tried to search the website of 100 largest hospitals in the U.S and called 50, but got responded by hospitals that they cannot release exact cost which may vary case by case.

It is urgent to develop easy and quick detection method given the significance of risks posed by antibiotic resistant pathogens. Research on the current status of containment strategies in the US and Korea make us reaffirmed that our project is worthwhile efforts to be pursued enthusiastically. Our team at Elan Vital Korea is drawn by the idea that we may be able to rapidly and efficiently detect infection by antibiotics-resistant bacteria using a synthetic biology methodology, quorum sensing (QS).

The White House, NATIONAL STRATEGY FOR COMBATING ANTIBIOTICRESISTANT BACTERIA, NATIONAL STRATEGY FOR COMBATING ANTIBIOTICRESISTANT BACTERIA

The White House, Executive Order -- Combating Antibiotic-Resistant Bacteria

Center for Disease Control and Prevention http://www.cdc.gov/

Korean Center for Disease Control and Prevention (http://www.cdc.go.kr/search/sEngine.jsp?kwd=MRSA%EA%B0%90%EC%97%BC_

Korea Antibiotic Resistance Monitoring System,(KARMS), Annual Report, 2009, 2011, Korea CDC, NIH
http://www.ncbi.nlm.nih.gov/pubmed/21140281

Hye Soo Lee, Hyun Lim, Evaluation of Various Methods for Detection of Methicillin Resistance Staphylococcus aureus (MRSA), Department of Laboratory Medicine, Chonbuk National University Medical School, and Institute for Medical Sciences, Chonju, Korea

Soo Jung Kim, Comparison Between Antimicrobial Susceptibility Test and mecA PCR Method for Reading of Methicillin-Resistant Staphylococcus aureus, Korea Journal of Microbiology, Vol. 47, 2011.12, p381-p385

Eun-Sung Moon , Paper Disc Method, MIC Test, CNU Biochemistry Lab.

R. STALONS and CLYDE THORNSBERRY , Broth-Dilution Method for Determining the Antibiotic Susceptibility of Anaerobic Bacteria Department of Microbiology, Health Sciences Center, Temple. University, Philadelphia, Pennsylvania 19140; and Antimicrobics Investigation Section, Center for Disease Control, Atlanta, Georgia 30333

Kyung Lee, Se Ran Heo, Soon He Choi, Sang Hoon Song, Kyoung Un Park, Junghan Song, and Eui Chong Kim, Comparison of Various Methods for Detection of Methicillin-Resistant Staphylococcus aureus Department of Laboratory Medicine, Seoul National University Bundang Hospital, Gyeonggi-do, Korea Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea

Jongyoun Yi, Eui-Chong Kim Microbiological Characteristics of Methicillin-resistant Staphylococcus aureus 1 Department of Laboratory Medicine, Seoul National University Hospital, 2 Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea

Hong-Bin Kim, Chong Moon Sa, Jaeil Yoo, Bong Su Kim, Ok Jin Yun , Hye Ryoung Yoon and Yeong Seon Lee, Antibiotic Resistance Patterns of Staphylococcus aureus Isolated from the Patients Admitted to Non-tertiary Hospitals Laboratory of Nosocomial Pathogens, Department of Microbiology, National Institute of Health (NIH) and Seoul Clinical Laboratories (SCL)* , Seoul, Korea

Kwon, Young-Il, Tae-Woon Kim , Hae-Yeong Kim , Yun Hee Chang , Hyo-Sun Kwak , Gun-Jo Woo , and Yun-Hee Chung,Monitoring of Methicillin Resistant Staphylococcus aureus from Medical Environment in Korea. Test and Research Center, Korea Consumer Protection Board, Seoul 137-700, 1 Institute of Life Sciences & Resources, Kyung Hee University, Suwon 449-701, Korea, 2 Department of Food and Nutrition, Myongji University, Yongin 449-728, Korea, 3 Center for Food Safety Evaluation, Korea Food and Drug Administration, Seoul 122-704, Korea

 Mayo Clinic www.mayoclinic.org/

 Massachusetts General Hospital www.massgeneral.org/

 Cleveland Clinic http://my.clevelandclinic.org/

 UCSF Medical Center www.ucsfhealth.org/

 Johns Hopkins Hospital http://www.hopkinsmedicine.org/the_johns_hopkins_hospital/

 New York-Presbyterian University Hospital of Columbia and Cornell http://www.newyorkpresbyterianhospital.com/

 Northwestern Memorial Hospital http://www.nm.org/location/northwestern-memorial-hospital

 Yale-New Haven Hospital www.ynhh.org/

 Asan Medical Center http://www.amc.seoul.kr/asan/main.do

 Samsung Medical Center http://www.samsunghospital.com/home/main/index.do

 Seoul National University Hospital http://www.snuh.org/

 Severance Hospital http://sev.iseverance.com/

 The Catholic University Korea Seoul Saint Mary’s Hospital http://www.cmcseoul.or.kr/

 Gachon University Kil Medical Center http://www.gilhospital.com/

 Korea University Medical Center http://www.kumc.or.kr/main/index.do

 Ajou University Hospital http://hosp.ajoumc.or.kr/