RAPID point-of-care infection detection and antibiotic-resistance TESTing enabled with laser-patterned microfluidic devices (RAPID-TEST)

Lead Research Organisation: University of Southampton
Department Name: Optoelectronics Research Centre (ORC)

Abstract

The aim of this proposal is to use our proprietary laser printing technique to develop low-cost, paper-based diagnostic tests addressing a genuine unmet healthcare need, namely the rapid identification of bacterial infections and antibiotic susceptibility profiles to guide therapeutic decision making. Our point-of-care (POC) tests will be suitable for use in a clinic or by the patients themselves as part of a pre-consultation screening process or on-going home monitoring.

We have already developed our paper testing platform and demonstrated excellent microfluidic properties, establishing the proof of principle that the devices can be impregnated with a colorimetric system capable of detecting the inflammatory marker, C-reactive protein. We are now building on this concept for developing excitingly ambitious and conceptually novel diagnostic tests for use at the POC. Our target diseases, urinary tract infections (UTIs) and the chronic airway disease in cystic fibrosis (CF) are in themselves serious and currently sub-optimally managed, but importantly, the successful technology will also be applicable to a large number of other medical and industrial applications.

Bacterial infections affect large numbers of people, with significant quality of life and healthcare cost consequences. There have been very few new antibiotic agents developed over the last 1-2 decades and there is increasing concern over the global epidemic of antimicrobial resistance (AMR). A major part of this problem relates directly to the widespread and indiscriminate use of broad-spectrum, non-targeted antibiotics. Choosing the correct antibiotic is however difficult in the absence of an accurate diagnosis. Current protocols for the identification of an infecting pathogen and follow-on testing of its susceptibility to antibiotics are time-consuming and require specialist microbiology culture-based procedures. These approaches are not only costly and inconvenient, but there is a period of diagnostic uncertainty during which treatment is chosen empirically and may be sub-optimal. Patients' health and well-being is adversely impacted and these delays contribute to the emergence of AMR.

Our proposed novel microfluidics-based devices will uniquely serve a dual purpose - first, rapid, POC identification of a pathogen and second, cheap and expedited testing for its antibiotic resistance profile. We will achieve this through the use of an optimised enzyme-linked immunosorbent assay (ELISA) to produce an immediate colour change on contact with the chosen bacterial antigen. We have selected three problematic bacteria causing UTIs and serious infection in patients with CF. Once single detection systems have been optimised, the next task will be to multiplex them onto the same device. Coupled with the detection system will be chromogenic, agar-based culture wells containing various antibiotics. After a short period of incubation, ~ 24 hours, colour changes will indicate the antibiotics to which the bacteria are resistant. This process would significantly reduce the current diagnostic time of 3-4 days.

These tools will be important and timely for GPs/Consultants in delivering an accurate antibiotic treatment of their patient's infections with significant savings in healthcare costs. For the patients, there will be reduction in symptoms and consequent improvements in quality of life. In the context of CF, a life-long disease, the devices could be used by patients in their own homes for long-term surveillance, in a fashion very aligned with the UK CF Trust's flagship SmartCare programme; this will not only empower patients in self-management, and facilitate earlier treatment, but we may ultimately be able to allow non-infected patients to have contact with each other - lack of real-time knowledge of an individual's infection status currently mandates strict segregation which has a negative impact on patients' well-being.

Planned Impact

Rapid point-of-care (POC) diagnostics has a crucial role both in developed and developing countries, due to the increasing strain on the public health system and the lack of resources. Use of on-site POC diagnostics is emerging as a popular choice for non-invasive testing for infections. Rapid identification of pathogens causing such infections is difficult to accomplish, and therefore the current routine practice is initial use of empirical antibiotic treatment which leads to antimicrobial resistance (AMR).

Economic Impact: The UK continues to experience an economic challenge, where large budget cuts due to austerity measures coupled with an ever-increasing demand for better healthcare provision is leading to an unsustainable healthcare scenario. Early-stage testing for bacterial infections and their resistance to antibiotics at the POC, a route proposed via our proposal, will enable crucial savings to the NHS (in terms of equipment, medicine costs and staff) thus shrinking the burden on hospitals via reduced patient trips to A&E units, emergency call outs and hospital admissions that alleviate hospital bed shortages.

Tests developed within the framework of our proposal which enable early-stage testing for infections, such as urinary tract infections which are the most common healthcare associated infections (HCAI) accounting for 17% of all HCAIs, cost approximately £170 million/year in England and constitutes up to 3% of all GP primary care consultations and for those patients needing hospital admissions, extending the average length of hospital stay by 6 days, accounting for an extra 800,000 bed days annually.

Such transformative screening diagnostic tools are being sought after by medical specialists, and this is the motivation of the proposed research, which will therefore translate into huge financial savings for the UK's public health system and its economy.

The methodologies proposed will make possible the large-scale manufacture of such paper-based POCtests, which are affordable and therefore would be widely adopted by the NHS and similar organisations abroad. The UK stands to make substantial gains therefore from the economic benefits that would ensue.

Societal Impact: As stated in the O'Neill report, diagnostic tests (such as those proposed by us) should be an important measure (Intervention no. 5) that must be adopted urgently to reduce the demand for antimicrobials, essentially antibiotics, and thus tackle the problem of AMR. To support this, governments, regulators and other health system leaders should consider incentives to facilitate the uptake and use of rapid POC diagnostics in primary and secondary care because they can also facilitate prompt exchange of invaluable clinical information between healthcare professionals and their patients from the comforts of their homes, which in some cases could be in inaccessible locations. Such remote testing removes the possibility of contracting infections from hospital visits. Since a timely exchange of clinical data could potentially be very useful in early detection and prevention of life-threatening infections, diseases or conditions, the wide societal benefits would not just be in saving lives but also improving the quality of life.

These rapid, dipstick-like tests, which lead to early detection of bacterial infections and testing for antibiotic-susceptibility, will facilitate prompt, accurate antibiotic prescriptions, potentially transforming treatments of millions of patients and minimising the threat of AMR. The tests are designed to be affordable and easy to use, becoming a routine part of primary and secondary care, outpatient settings and hospital screening for all patients. Furthermore, the applicability of low-cost, disposable paper-based testing developed via this project could also be extended to include testing for screening of food pathogens and environmental toxins, all of which inarguably have massive societal benefits.
 
Description We have been successful in growing our targeted infection-causing bacteria (e.g. E-Coli and Pseudomonas Aeruginosa) on paper-based devices, which have been previously impregnated with agars (i.e. a bacterial growth media), specific to the different bacteria. We have identified the correct parameters (volumes, concentrations etc.) required to grow both E. Coli and Pseudomonas. Additionally, we have tested for a range of different antibiotics that can inhibit growth of such bacteria. We have incorporated these antibiotics in our paper-based devices, and we have successfully identified the correct parameters (volumes, concentrations etc.) that would allow us to differentiate between resistant and susceptible bacterial pathogens. Finally, we have identified sets of antibodies that are specific to E. Coli and Pseudomonas, and have developed a lateral flow device that would detect these bacteria. We have yet only performed some initial experiments and further development and optimisation are underway. We believe we are on the correct route and on target for the manufacturing and testing (using real patient samples) a complete device (that uses agar-impregnated zones and a lateral flow geometry) for detection and antibiotic susceptibility testing of the mentioned bacteria.
Exploitation Route The early-stage results relating to the above described key-finding has been reported at conferences and the details of these are available for reading via the relevant conference proceedings listed on the University of Southampton's openly accessible database (Pure) stated above. As part of the award we will further explore these outcomes and further undertake a detailed study, however, it can, if desired taken up by an healthcare diagnostics manufacturer to produce low-cost bacterial detection tests. To explore the usefulness of these early results we have engaged with Public Health England, Porton Down, who are interested in exploiting these early results for their screening of E-Coli-related infections. We are also in contact with several commercial companies that have been very interested in our progress, and would be willing in the future to adopt our technology.
Sectors Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

URL https://pure.soton.ac.uk/admin/workspace/personal/family/researchoutput/
 
Description Partnership with Public Health England at Porton Down 
Organisation Public Health England
Department Public Health England Porton Down
Country United Kingdom 
Sector Public 
PI Contribution We have generated some early results that indicate the possible usefulness of laser-patterned, and growth-media impregnated paper tests that could be possible tools for early-screening of bacteria causing Urinary Tract Infections and Upper Respiratory Tract Infections.
Collaborator Contribution Public Health England are interested in using this methodology for screening of E-Coli related infections and are keen to provide inputs in the form of their valuable expertise and furthermore have also shown willingness to allow access to their facilities to enable testing of our paper-tests.
Impact Yes, this is multidisciplinary collaboration between laser-physicists or engineers at the University of Southampton and biologists at Public Health England providing the complementary skills much needed to assist with the work pursued via this award at the University of Southampton.
Start Year 2017
 
Description Southampton General Hospital - Pathology Laboratory 
Organisation Southampton General Hospital
Country United Kingdom 
Sector Hospitals 
PI Contribution As part of this funded research, our team at the University of Southampton are developing tests that allow for detection of bacterial pathogens responsible for Urinary Tract Infections. As described below, as part of this research collaboration, we would test these devices within the laboratory of our collaborating organisation - Southampton General Hospital.
Collaborator Contribution The tests that we have been developing at the University of Southampton via this funding would be tested at the Pathology laboratories of the Southampton General Hospital. The partnering team at the laboratory would assist with the testing, and more importantly, would make available patient samples for the pre-clinical testing. The results and further discussions with pathology lab team would enable us to assess the performance of our test and would also help us understand if any alterations or modification are desired for our initial prototype. As this work would be done with professionals, who are engaged in such testing regularly, this would also allow us to refine and tailor our tests as per the real-world needs.
Impact We are in the process of tetsing the devices and the outcomes would happen later in the year.
Start Year 2018
 
Description 2nd Global-NAMRIP in Kampala, Uganda 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr Collin Sones the PI of the award attended and presented the work funded via this award relating to use of low-cost paper-tests for identifying bacterial pathogens at the Global NAMRIP 2019 conference in Kampala, Uganda. The purpose of the meeting organised by the University of Southampton's Network for Anti-Microbial Resistance and Infection Prevention (NAMRIP)(https://www.southampton.ac.uk/namrip/index.page) was to engage with the healthcare professional, policymakers and academic stakeholders to understand and tackle the problem of antimicrobial resistance acorss the African continent. The meeting was attended by more than 100 people from various goverment and private organisations/institutes that worked in human and animal health, sanitation, water management and social sciences, and Dr Sones and other members of the NAMRIP/University of Southampton showcased their research as possible solutions for tacking this global challenge.
The PI Dr Sones has now made valuable contacts with the Ugandan government's national health laboratory, Uganda National Health Services (UNHLS) in Kampala, who are keen utilise the paper-based, point-of-care testing technology developed for identifying bacterial pathogens for other bacterial pathogens namely, MRSA and ESBLs, which are a major health concern in Uganda.
This engagement would provide a useful collaborative pathway that validates the paper-testing technology developed via this funding and would also provide a route to extend its application domain in to testing of other pathogenic bacteria.
Year(s) Of Engagement Activity 2019
URL https://www.southampton.ac.uk/namrip/news/2019/02/uganda-conference.page?
 
Description Global NAMRIP 2018 - Ghana 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr Collin Sones the PI of the award attended and presented the work funded via this award relating to use of low-cost paper-tests for diagnostic testing at the Global NAMRIP 2018 conference in Accra, Ghana. The purpose of the meeting organised by the University of Southampton's Network for Anti-Microbial Resistance and Infection Prevention (NAMRIP)(https://www.southampton.ac.uk/namrip/index.page) was to engage with the healthcare professional, policymakers and academic stakeholders to understand and tackle the problem of antimicrobial resistance not only in Ghana but throughout the African continent. The meeting was attended by 30 such people from various organisations and Dr Sones and other members of the NAMRIP/University of Southampton showcased their research as possible solutions for tacking this global challenge.
Year(s) Of Engagement Activity 2018
URL https://twitter.com/search?f=tweets&q=collin%20sones&src=typd
 
Description NAMRIP - Summer Conference 2017 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Prof R W Eason (Co-I) and Dr Peijun He (PDRA) attended and presented the initial ideas and results relating to this award at the NAMRIP summer conference 2017 which had a very distinguished speaker, Lord O'Neill. The conference titled, 'Bridging the Gap between Engineering and non-Engineering research to fight AntiMicrobial Resistance' showcased the various research pump priming projects that were funded by NAMRIP, and had the common objective of fighting the menace of Antimicrobial Resistance. The work presented included that relating to changing behaviour in healthcare settings to how laser printing can make a paper based low-cost diagnostic tool to combat TB and related bacterial infections. The latter is work arising for this award. The hands-on demo of colour changing paper-tests that identify infections was also showcased by our team and was a very simple visual way to communicate the usefulness of the work funded by this award - it engaged a vast audience very keen to learn about tools to tackle Antimicrobial Resistance.
Year(s) Of Engagement Activity 2017
URL https://www.southampton.ac.uk/namrip/news/events/2017/06/conference-2017.page?
 
Description NAMRIP at New Forest Show 2017 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact The PI Dr Sones participated as a public-outreach team-member of (the University of Southampton's strategic research group) Network for Anti-Microbial Resistance and Infection Prevention (NAMRIP), which demoed their exhibit - The Most Dangerous Game In The World at a regional show held annually, 'New Forest Show 2017'. This exhibit which is a collection of five games, arranged in the form of an arcade-style game was designed and made at the Winchester Science Centre, and its objective is to teach people, particularly children, about Antimicrobial Resistance (AMR) and safer use of antibiotics. Additionally, this also provides with an opportunity to educate the general public engaging with the game with the various AMR-related research activities undertaken by NAMRIP members. Through several conversation Dr Sones was able to talk with people from a wide audience about this award and the work on their use of low-cost paper tests for early-stage identification of common infections such as Urinary Tract Infections and Upper Respiratory Tract related bacterial infections in patients with Cystic Fibrosis.
Year(s) Of Engagement Activity 2017
URL https://www.southampton.ac.uk/namrip/news/2017/07/namrip-at-new-forest-show-2017.page
 
Description NESTA Longitude Prize, Antibiotic Awareness Week event - NAMRIP Public Outreach 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other audiences
Results and Impact The PI Dr Sones participated as a public-outreach team-member of (the University of Southampton's strategic research group) Network for Anti-Microbial Resistance and Infection Prevention (NAMRIP), which demoed their exhibit - The Most Dangerous Game In The World at 'NESTA Longitude Prize, Antibiotic Awareness Week event'. This exhibit which is a collection of five games, arranged in the form of an arcade-style game was designed and made at the Winchester Science Centre, and its objective is to teach people, particularly children, about Antimicrobial Resistance (AMR) and safer use of antibiotics. Additionally, this also provides with an opportunity to educate the general public engaging with the game with the various AMR-related research activities undertaken by NAMRIP members. As an example, through once such conversation Dr Sones was able to talk with a participant from Medicine Sans Frontiers about this award and the work on use of low-cost paper tests to identify infections.
Year(s) Of Engagement Activity 2017
URL https://www.southampton.ac.uk/namrip/news/2017/11/nesta-event.page