PERP-ID - Point of care diagnostic for lung pathogens of pigs

Lung infections are responsible for substantial suffering, deaths, and economic loss in the worldwide pig industry. Infections are caused by bacteria, viruses or a combination of the two. Control of lung infections is by the use of good farm practices (e.g. avoiding overcrowding), antibiotics and/or vaccines. However, antimicrobial resistance is a problem and there is a lack of effective vaccines for most pathogens, hence, pig lung infections are a substantial burden to the industry. For effective control, identifying the pathogens causing pig lung infections is crucial and dependent on diagnostic tests. However, current tests are slow - it can take many days to get the results back from the laboratory - and many tests are required, one for each bacterium or virus. At the request of extensive feedback from veterinarians, pork producers, farm managers, and policy makers, we propose to develop one test that can rapidly and cheaply detect major bacterial and viral lung pathogens of pigs,and can be used at the farm - a so-called a point of care test. The PorcinE Respiratory Pathogen-IDentification (PERP-ID) device will be designed to identify the five bacterial and three major viral pathogens of pig lungs, and will be based on an electronic system developed at Imperial College London that can rapidly detect the presence of specific DNA molecules. Results can immediately be sent from the farm to local and/or global locations via a smart phone enabling fast remedial action at the farm e.g. giving appropriate antibiotics. Once developed, we will analyse clinical samples and compare the results obtained from PERP-ID with those from currently used diagnostic tests. The successful development of PERP-ID will allow more rapid and cheaper diagnosis of lung disease in pigs caused by bacteria and viruses, and result in a substantial reduction in suffering, death, antibiotic use, and economic losses in the worldwide pig industry.

The porcine respiratory disease complex (PRDC) is a substantial cause of morbidity, mortality, and economic loss in the worldwide pig industry. PRDC pathogens include bacteria i.e. Actinobacillus pleuropneumoniae, Haemophilus parasuis, Mycoplasma hyopneumoniae, Pasteurella multocida, and Streptococcus suis, and viruses i.e. porcine circovirus type 2, porcine reproductive and respiratory syndrome virus, and swine influenza virus. Diagnostics are crucial to PRDC control enabling surveillance, identification of emergent PRDC-associated agents, and the use of appropriate antimicrobials and/or vaccines. Typically, many tests are done (each one identifying a specific pathogen), and are laboratory-based, delaying the time to remedial action. This proposal is a direct response to extensive stakeholder feedback (pork producers, veterinarians, policy makers) requesting a rapid cheap point-of-care (POC) test to identify major pathogens of the PRDC. Building on results demonstrating feasibility with detection of carbapenem resistance in Klebsiella pneumoniae and zika virus, we plan to use a platform that combines isothermal amplification of DNA (i.e. at a constant temperature) coupled with detection by complementary metal-oxide semiconductor (CMOS) technology, that can detect the aforementioned bacterial and viral pathogens of the PRDC. The PorcinE Respiratory Pathogen IDentification (PERP-ID) device will be directly compared with conventional laboratory-based diagnostics. To our knowledge, this will be the first single POC (on farm) test that identifies major PRDC pathogens. Additionally, PERP-ID will allow rapid dissemination of results locally and globally via a smart phone, enabling fast remedial action. The successful development of PERP-ID will allow rapid, cheap diagnosis of PRDC, and result in a substantial reduction in morbidity, mortality, antibiotic use, and economic losses in the worldwide pig industry.

The porcine respiratory disease complex (PRDC) is a substantial cause of morbidity, mortality, and economic loss in the worldwide pig industry. Crucial to the success of PRDC control is the availability of diagnostics to enable surveillance, identification of emergent PRDC-associated agents, and the use of appropriate antimicrobials and/or vaccines. We plan to develop a single cheap rapid diagnostic platform called the PorcinE Respiratory Pathogen-IDentification (PERP-ID) device that detects the major five bacterial and three viral pathogens of the PRDC.

Success would offer a step change in capability for combating PRDC, but would have far reaching impact for the diagnosis and treatment of other veterinary and human bacterial diseases more broadly. This work closely addresses the key BBSRC Strategic Priorities of 'Animal Health', 'Combating Antimicrobial Resistance' and 'Welfare of Managed Animals', and the beneficiaries of this research and how they will benefit are as follows:

*Academic Researchers: through broad academic advancement and instigation of new research programmes
The data generated will be of immediate benefit to researchers in Universities, Research Institutes and Industry with interests in diagnostics, epidemiology, population biology, pathogen-pathogen (polymicrobial infection) and host-pathogen interactive biology research more broadly. Focusing on the immediate beneficiaries, these include, among others, scientists in the UK and abroad working directly on PRDC and pig respiratory disease. Once PERP-ID is validated, an immediate avenue of investigation will be to confirm whether the approach has broader applicability to other veterinary and human infections, leading to a potentially rapid evolution of related diagnostic devices.

* The veterinary commercial sector: through exploitation and commercialisation
Our research aims to provide the basis for a diagnostic device that will be readily exploitable by the UK pig industry. This is an exemplar project. Success will facilitate translation to point of care diagnostic development for infectious diseases of other animals (cattle, sheep, chickens, dogs and cats). In the longer term, a successful conclusion of this (and future related) projects and introduction to the market will create wealth and economic prosperity through increased turn-over, profit and exports, and creating and safeguarding jobs for the company and workers involved.

* The pig industry: through the ability to rapidly diagnose PRDC
In the longer term, successful translation of PERP-ID will reduce: medication costs, pig suffering and mortality and the significant economic impact of PRDC on the swine industry. In particular, rapid diagnosis will lead to a reduction in the use of antibiotics by preventing their use for viral and allowing selection of appropriate antibiotics for bacterial pathogens. Thus there will be a positive impact on antimicrobial resistance, a recognized global threat.

* The general public: through increasing public engagement with research.
Throughout it will be possible to engage with the general public and local schools to raise the profile of the research area and project. This will encourage interest in biosciences research and UK innovation and provide a context to local bioscience education.

* Researchers working directly on the project: thereby contributing to a highly trained workforce.
The skills gained by the researchers working on this innovative interdisciplinary project will be highly transferable; providing excellent training to enable them to embark on a successful career in any setting including academia or industry. For example, working on a joint academia-industrial project provides experience in collaboration and team working, communicating effectively with non-specialists, and via publishing papers, patents, and conference presentations they will hone their verbal and written skills.