Hi, my name's Fleur Whitlock, and I'm a vet at eququine Infectious Disease Surveillance, otherwise known as EIDS, and I'm talking here today about why diagnostics matter. So zooming in on how we can improve equine outcomes utilising rapid respiratory syndromic testing such as the one offered by Baya Marie. Just to briefly introduce first the group that I work for, so EIDS are based at the University of Cambridge and we're generously supported by the UK Thoroughbred racing and breeding industry.
And really the aims of what we do are to control and prevent equine infectious disease in our UK population. We primarily focus on endemic disease, but we do work closely with government for those diseases that are that are under legislation as well, so notifiable diseases. I just want to introduce a concept of what we rely on to conduct a lot of our surveillance, and this is the surveillance wheel pathway.
And you'll see around the outside various different stakeholders and different colours who we rely on to have this wheel turning. And we'll come back to this concept as we go through the next 15 minutes or so. But just to introduce how this works, we're reliant on an owner realising their horse is sick, and being aware of the clinical signs of infectious disease, contacting their vet in a timely manner, and that vet visiting that case and taking the correct samples from that animal for the potential differentials it may be suffering from.
And then either conducting in-house testing or submitting that sample to a laboratory. And then reliant on if you do get a positive result, sharing that with ourselves at EIDS so we can have information on how that particular pathogen is behaving in the population that it's infecting and also on a molecular level how that pathogen is looking too. So it's quite a long arm that we rely on to get that surveillance wheel turning and encourage you to think about how you can work with us to get that wheel turning.
We have various different initiatives in place to encourage engagement with the work that we do. We have some free testing initiatives for flu, to encourage uptake at that section of the surveillance wheel. And really importantly is sharing information across the industry and keeping people aware of what is going on out there in the world of infectious disease.
So we have a number of different real time reporting systems. The Telltale text message alert System is a specific look at UK occurrences of flu, herpes viruses when they're in the neurological abortion or neonatal death presentations and also notifiable diseases too. It's a really.
Encourage you to sign up for that. And we also oversee the international reporting of disease through the International collating Systems of various different groups like ourselves based in the UK and we'll also be based in other regions, and they will report their own infectious disease outbreaks and then we put them all together and almost daily if you're signed up for this, you'll get a real-time report via email from the ICC of the occurrence of infectious disease internationally. We also have some specific surveillance initiatives for strangles and equine flu too.
So on the left you can see a tick list of how you as an equine vet can get involved in the work that we do and how you can sign up for the various different initiatives that are available and I'll share a QR code at the end that will take you through to the sign up page for you to be able to get involved in these. So why we're here today, why do diagnostics matter? Firstly just to have a think about a concept of.
Various different phases when you're dealing with an outbreak. So we've got detection, identification of disease in a population, going back to the surveillance wheel, that's an owner obviously noting their horse is sick, contacting you, and then you doing some diagnostic testing. We've then got the containment phase where we're really trying to stop that fire burning on that population and also controlling disease on that within that population and not, not letting it go beyond to the wider population.
And then considering how we can obtain clearance of disease as well. So firstly, diagnostics really do underpin that effective outbreak response and hopefully in the next few slides we'll sell that to you as to why that is. So in terms of detection, we need to conduct testing to understand what pathogen are we dealing with, and that needs to be fast and accurate and systems like the one offered by B Amiru really do offer that.
We need to consider what pathogen we're dealing with as that's going to really determine what outbreak response approach we're gonna take for that population. Our various different endemic infectious respiratory diseases in the UK are quite different in terms of their epidemiology, so thinking about how they transmit amongst populations and how we're gonna stop them spreading, it's really key and without knowing which one we're dealing with, we won't really know how to conduct our outbreak response. And then finally to really hammer home, clinical signs alone cannot identify the pathogen, and I'll go on to why this is as well, but just bear that in mind, they can all look pretty similar.
Containment is consideration of how can we contain disease within that population, stopping its spread within that population too that we're dealing with. So generally this will be a single premises, so encouraging the immediate isolation of cases. So a lot of the time it may be if you're sending a sample to the lab, you're gonna be waiting for the postal system, you know, it could be a weekend in the way as well before you get your results.
And trying to encourage a client. Isolated case and we don't know if they have an infectious process, although they're presenting with signs that are very indicative of. It can be very hard to encourage clients to instigate those sorts of measures.
So actually points that these point of care tests or desktop testing systems enable us to have a little bit more welly behind encouraging clients to instigate these sorts of measures. We'll hopefully have a result within a couple of hours and we can get back to our practise in time. Encouraging the premises to introduce voluntary movement restrictions as well can be really challenging and really hard for us to communicate, but the benefits of the wider for the wider population are just really high, so we do need to encourage that.
And again, having a diagnostic test result might give you a little bit more ability to get them to do so. And then we really should be encouraging heightened biosecurity measures amongst that population to enable containment. So definitely temperature checking of.
Obviously the first thing an animal's gonna do when it spikes when it gets an infectious process is spiking a fever. So twice daily temperature checking and recording those is really integral and totally free. And so we really do encourage clients to do that.
And then potentially other measures might include things such as vaccination, but obviously that would be very pathogen specific, and again we need the diagnostic test result to understand what pathogen we're dealing with, to understand how best we can conduct our containment. Control is really thinking around this risk based zoning system that I think you all will be familiar with, so that's the red red, amber green traffic light system. So segregating groups based on whether they're currently infectious, so that would be the red group.
Amber, so those that have had known contact with the red horse, and then green horses where they've had no known direct or indirect contact. So a lot of premises may not have a green group, but utilising laboratory testing to understand how you're going to designate these risk-based groups can be really helpful for particular pathogens. That's really gonna aid our evidence-based decision making.
It might not be something we do so much for presentations of flu, but if we're dealing with EHB1 neurological disease. Or strangles utilising diagnostic testing for our control is integral to the work that we do at EIDS when we're aiding vets in outbreaks. And finally, clearance and the requirement for testing to aid clearance decisions will really depend on the pathogen that we're dealing with and the particular outbreak scenario.
Again for things like strangles where you've got a carrier status. And EHV one neurological presentations where the rest of the population might be relatively subclinical but still potentially shedding and an infectious risk, diagnostic testing is integral. Again, depending on the number you're testing, you may have to make a decision as to whether desktop testing systems are the correct ones to be using in that scenario or whether you need to be outsourcing that to a lab.
But testing in for clearance is is really important in certain circumstances. So consideration around how we can use our case history and how that really is the foundation for when we're making diagnostic decisions. So how, how are we going to use our case history and what's that going to do?
So it really does influence who we're going to sample, so understanding when you've got a horse presented in front of you. Is that the best horse to be sampling? We're relying on agent detection testing modalities a lot of the time, so we, we want the infectious agent to be at the sites that we're sampling.
So for endemic respiratory diseases, that's going to predominantly be the nasopharynx. You may be doing, guttural pouch lavage potentially, but just bearing in mind if strangles is. Top differential, it's gonna take around 2 weeks potentially for the pathogen to make its way to the gut guttural pouches for you to find it there from, exposure.
So just bear that in mind. You can also obviously swab abscesses and also take, aspirates of abscesses too, but predominantly it's going to be nasopharyngeal swabbing. So a lot of the time you'll be being called out to see a particular case by your clients, may be you've got multiple different vets on that premises or vet practises, so you are limited to just that horse in front of you.
But just bearing in mind when you're getting your history, trying to understand how long has this animal been sick for. And if we go back to that surveillance, where a lot of the time unfortunately plants are ringing us when we're already potentially 7 days into the onset of disease disease process in that animal. So it may be the pathogen's been and gone from the from that animal when you come to sample it.
So just bear that in mind when making sampling decisions. What samples to collect, as I've said, it's predominantly gonna be nasopharyngeal swabs, but consideration of what other samples we may take to, and run on systems such as this. And then differentials to test for.
Obviously there are 3 endemic respiratory diseases in the UK that will be the primary pathogens that we'll be concerned about. And as I said, they may all look the same, and why is this? They might have had prior exposure amongst the population.
You might have a population that's vaccinated or even a subset that's obviously going to lessen the severity of clinical signs. It can also be very pathogen strain dependent too and depending on the scenario. So these are just some of the variables as to why these infectious processes may all look the same, particularly in that acute phase.
You might just have a pyrexic horse that's slightly off its feed with a slight snotty nose, so don't rule any of them out at that stage. Classically, if we're kind of being patho pneumonic with how these present, equine flu generally is very contagious, so you get fast spreading signs amongst a naive group, but potentially your group may not be fully naive, and it's this classic kind of hacking cough that you hear, it may not be. Equine herpes virus one, obviously you can see respiratory signs, a snotty nose, slight cough, .
They may have a subset of the population with neurological signs, and you can get outbreaks where you get mixed infections with all three presentations and abortions too. Equine herpes virus 4 predominantly will be respiratory signs. We will never see an abortion storm due to EHV 4, it would be a single case if, there was a cause of abortion, and we're yet to be convinced by a case of neurological disease caused by EHV4.
So the predominance of EHV4 can actually be quite subclinical or very mild respiratory disease. Strangles and where it gets its name from, so causing that lymph node abscessation that effectively, effectively strangles the horse and and makes it very tricky to breathe, and we don't see those presentations so much anymore. So they can be pretty subtle, they may have no signs of just a very, very slight snotty nose, so don't be caught out by those.
I'm so hoping you'll realise where I'm going with this, the best strategy you can have. Is to run a full respiratory pathogen panel for these to make sure you're covering all bases. And just bear in mind as well, on occasion you may getcur infections, particularly the herpes viruses coming along to the party if an animal's been confirmed to have, flu or strangles, particularly EHV4 you may find doing that, so it's understanding the clinical relevance of that scenario.
And then finally, when we're thinking back to case history as a foundation, diagnostic, it's going to really influence your diagnostic result interpretation. So going back to the history and thinking about can I trust these negative results that I've gotten, it can be really frustrating when you get a bunch of negative. Results, but can you trust those?
Did you sample the animal at the right time? Did you sample the right animal? Did you take the right samples?
Were you happy with your sample? It may be you need to go back out and sample some others. And again, if you have, desktop testing in your practise, that's a really easy thing for you to be doing.
So don't rule out, the endemic diseases that we have testing available for just based on one single negative swab from one animal. And we're here to help at EIDS. We really encourage you to engage with us.
We will, you know, we, we've got a free veterinary advice service and we can assist you every stage of an outbreak, response. So through from stable side diagnostic testing, deciding what samples to take from what animals, how you can control disease and contain it to a premises, thinking about. How or if we need to conduct any clearance and then most importantly, how can we prevent disease in that particular population in the future.
So we can have a pragmatic approach considering the various risk factors for that population and come up with a plan altogether and we're always here to help. So please do save our number in your phone and give us a call, we'd be really keen to hear from you. So how can you contribute to surveillance, thinking back to that surveillance wheel and how important people that are conducting desktop testing will be to contributing to surveillance too.
We've got lots of labs that contribute to the work that we do and we will really will be reliant on vets in practise to do so too if you're utilising these systems. And so we have a website for reporting epidemiological data when you're dealing with an outbreak. I'll talk through what information is shared through that system, and how you can do so.
And then really importantly as well, if you have a sample that is positive, sharing that with our isolate library that's based at Rossdell's Laboratories in Newmarket is really integral. If you send us a flu sample, that'll be sent on to virologists at the University of Cambridge. And they conduct whole genome sequencing on those samples in near real time so we can understand how that flu virus from your particular infectious horse has changed based on the previous ones that we've seen over time and will current vaccines impart a level of protection that we expect them to do so, so really important.
And then herpes virus samples will be shared and stored in the freezer for future testing. And we also do conduct surveillance of strangles, that's through a slightly different arm, this SES system that I presented on one of my first slides. You can again share epidemiological data to SES and.
Similar way to our web form and the a team member that oversees that can inform you on how you can subscribe to SES. Unfortunately we're not conducting any molecular testing currently on SEqui positive samples, but we have done in the past and produced some really cool research out of it. So what happens to the outbreak data that you share?
So you go on the website, . If you have a positive case of flu or herpes presentations, and that will then log the data into an encrypted database and the access to that is restricted to the EIDS team. You have to, confirm that you're happy for, anonymized reporting to occur through the system, and if you do so, we only ever report at a county level, and here is an example of one of the reports we may share.
So we discuss the dates that the lab confirmed the case, some background history on the case, whether it's vaccinated or not, it's age, and whether it's a thoroughbred or a non thoroughbred. Any particular risk factors that have occurred, so a recent new arrival, clinical signs that it's demonstrated, and then how the positive diagnosis was made, and then we like to also acknowledge the laboratory that conducted the testing or the veteran practise, if it's appropriate to do so. So where is this information shared, as I've introduced the telltale text message alert system that you can sign up if you're a UK vet.
We'll share information on that particular outbreak for awareness weight raising and being, you know, aware of how are these pathogens behaving in our populations. What were the particular risk factors for this scenario, for example, if we're suddenly seeing a cluster of flu cases in vaccinated animals, that may make us think, well, we need to have a look at that virus and see what's going on with it, why a vaccine is not imparting the level of response that, that we should see. So really key to engage with surveillance.
Also, the international system that I introduced earlier, the ICC, will also have a very similar link shared, through its real-time email alert system. So final slide from me. I'm hopefully I've managed to convince you why using diagnostics will result in better outcomes for your horses, your clients, and the wider equine industry.
The importance of case history and utilising rapid testing, enabling you to make informed decision make informed decisions when dealing with an outbreak. We're here to support you at EIDS and really pleased you get in touch with us, we'd really love to hear from you. And then surveillance is only gonna work if we all contribute to it, and we're really passionate to encourage you to do so.
So in the top right corner you'll see a QR code that you can scan on your phones that will take you through to our website. We've got a sign up page on our website, for the various initiatives I've discussed today. And we've also got various different social media platforms you can, follow as well.
Many thanks. Hello, everyone. My name is Frederick Leon.
I'm an associate director of R&D at BMRe based in Greneble in France. So I have two decades of experience in molecular biology and microbioecology focusing on developing, innovating in vitro diagnostic solutions. And I'm very pleased to be part of this webinar and to share how BMRA can contribute to improving equine outcomes through the rapid respiratory syndromic testing.
So a few words regarding Biomeio. Bio is a family-led company which has grown to be a world leader in in vitro diagnostics, paving the way to help make the world a healthier place. So for over 60 years, Bioia provides diagnostic solutions, including systems, reagents, software, and services that determine the origin of a disease to improve the health of patients.
So today, Bioio is present in 45 countries and employs the most 15 people, 15,000 people. To address public health challenges, BORU invests 12% of its annual revenue in R&D, driving innovation in diagnostic solutions. So these efforts have made BioMieux a global leader in molecular syndromic testing.
Within hospitals and have paved the way for the development of innovative PSA point of care solutions. So today, BMaria brings its proven human health technology to veterinary diagnostics because pathogens know no borders between species and neither does diagnostics. So as Fleur explained, we believe that the location of testing is a key factor in diagnostic impact.
When discussing sample shipments to external labs with veterinarians, they often mention delays, logistical burden, or even sample degradation, and all of which can compromise results quality and slow down decision making. So we are therefore convinced that performing PCR testing directly in the clinic offers faster turnaround times, enabling quicker biosecurity actions, better clinical decisions, and potentially improved treatment outcomes. It also gives veterinarians greater control over the diagnostic process, allowing them to communicate results more quickly and clearly to horse owners, building trust and improving satisfaction.
So as we have seen, internalising PCR testing in the clinic makes a lot of sense, but that's only the beginning. So as a, as a leader of the veterinary R&D molecular team in Lee Marie. The challenge of my team is to develop a solution that truly meets, clinical needs.
So what are the main challenges we face in this development process? The first one is regarding clinical relevance, because clinical symptoms are often non-specific, veterinarians cannot reliably choose to test for just one or two pathogens, so this increases the risk of having to repeat tests, losing valuable time and potentially rising costs for the animal owner. So as I mentioned, a syndromic approach is essential, but then comes the question which pathogens should we include?
Ideally, we focus on primary pathogens, those whose detection provides meaningful information and helps veterinarians make confident clinical decisions. Opportunistic pathogens, while sometimes relevant, are often secondary, and their presence alone doesn't support clear decision making. The second point is regarding workflow, simplicity and safety.
So once the sample is collected from the horse, it's essential to use. The simplest possible method to introduce it into the PCR process. Ideally, a one-step piping technique should be preferred to minimise the human error and maximise the chance of obtaining reliable results.
And similarly, using a closed PCR system where tubes stay sealed during the entire process is essential to prevent contamination. During amplification, a single positive sample can produce billions of DNA copies which, if released into the lab environment, could lead to false positives in future tests. So the last point is regarding technical performance.
So beyond the practical aspect of user handing, it's essential that the technology we offer matches the sensitivity, specificity, and robustness of external reference laboratory tests. So to achieve this, we must be able to laze even the most resistant microorganisms and efficiently extract and purify very small amounts of nucleic acids from complex sample matrices. PCRSAs also need to be precisely designed to ensure high sensitivity and specificity for the targeted pathogens, and of course the icing on the cake is delivering results as quickly as possible.
So it's important to know that all these challenges were also identified and addressed during the development of our point of care molecular solution for human health, resulting in the launch of the powerful BioFrepofire system. So the Spotfire system is a compact, scalable, and easy to use platform, delivers rapid, reliable results in under 20 minutes with minimal hands on time. So with just one sample and one disposable cartridge, the system detects multiple pathogens in parallel, all associated with a specific nickel syndrome, such as respiratory infections, and provides a clear, easy to read report.
So we therefore made the decision to transfer this technology from hospitals and medical practises to veterinary clinics. So on the R&D side, our mission was to develop the first syndromic panel specifically designed for emergency situations in equal care. So let's take a closer look at the vet fire equine respiratory panel menu.
We decided to include the most relevant respiratory pathogens. Of course, the four most critical ones are EHV1, EHV 4, equine flu, and Streptococcus AKA. Additionally, The panel covers 3 benign viruses, equine rhinitis A and B viruses, and equine adenovirus 1, which are part of the differential diagnosis of infectious respiratory.
So as mentioned by Fleur, the key advantage of this syndromic panel is its ability to rapidly and reliably identify the causative agent from the very first test, eliminating the need for retesting and minimising the risk of misdiagnosis. So this approach also supports epidemiological surveillance because this systemic approach helps to monitor the true prevalence of circulating pathogens and can act as an early warning system. So now let's look at how simple and fast the testing process is with the Spotfire system.
So sample preparation is straightforward and takes only 2 minutes of hands-on time. First, you respond the equine nasopharyngeal swab in saline, then transfer 300 microliters of the sample into the fave using a pipette. Then add the sample buffer, which is a lysis buffer, draw the mix into the pouch, and load it into the Spotfire system, and that's it.
The system takes care of the rest, and this streamlined workflow is ideal for emergency situations where speed and reliability are essential. So when the sample is loaded, the spot fire system takes over with its unique labbing approach technology. So first, the samples move into the lysis chamber where be beating occurs.
So this mechanical lysis step use ceramic beads agitated at a high speed to break open all cells and microorganisms, even the most resistant ones such as RAM positive bacteria like Streptococcus echi. Then the released nucleic acids are bound by magnetic beads which are then moved from the lysis chamber to the purification chamber where a series of wash steps remove protein, cellular debris, and other potential inhibitors. The last step of the extraction process is the release of the purified nucleic acids from the base thanks to an emission buffer.
So each of these steps is essential to achieving high detection sensitivity, especially from complex matrices. So now this pure extract of nucleic acids moves to the first stage PCR chamber. A reverse transcriptor step is performed to convert any target RNA into DNA.
This is followed by a high order multiplex PCR involving dozens of primers which amplify the genomic material specific to each targeted analyte present in the sample. And this step is essential for improving sensitivity. Amplification products from the first stage PCR are then diluted to remove any remaining PCR primers and are combined with a fresh master mix and then allocated to each well of the array.
So each well of this array is precipotted with one pair of 2nd stage PCR primers. The 2nd stage PCR primers are designed to amplify sequences contained within the product of the 1st stage PCR. And this eliminates non-specific products associated with traditional multiplex PCCR because the second stage PCR specifically amplifies only target DNA.
And a double strand DNA binding dye is used to monitor each reaction. So after PCR 2, the system performs a high resolution DNA melting analysis and measures fluorescence in each well to detect PCR product. The software then checks the melting temperature and compares it to the expected range.
If the melting temperature matches, the result is positive. If not, it's negative. And each essay is tested in triplicate.
So to confirm a positive result, at least 2 of the 3 wells must be positive with similar melting temperature values, otherwise the result is negative. So this melt-based analysis greatly increases the specificity of the vet fire test, ensuring highly reliable results, and this entire workflow is completed in less than 18 minutes. So once the test is complete, the Spotfire system generates a clear and easy to interpret report.
As you can see on this slide, the report provides a summary of the test type, the result, if they are negative or positive, and a detailed list of detected pathogens. A check mark indicates a positive detection, and when needed, an action bar appears to guide interpretation. So this format ensures that even in high pressure situations, veterinarians can quickly understand the results and take appropriate action without needing to consult complex lab data.
So now a few words regarding the performance of the vet equine respiratory panel, and we can say it's not just fast and easy to use, it's also highly sensitive. The test can detect just a few dozen to a few 1000 copies of viral or bacterial genetic materials per millilitre of sample. This means even low level infections can be identified early, which is critical for timely intervention and preventing further spread.
So based on the limit of detection data available in the literature, our test demonstrates a higher analytical sensitivity compared to currently known methods. The panel has also undergone extensive inclusivity and exclusivity testing to support the reliability of the bed fire panel. So inclusivity has been evaluated in silicon and in vitro with over 76 viral and bacterial isolates, and this ensures that the test can detect a wide range of relevant strains and variants even at very low concentration.
Exclusivity testing has also been evaluated in silicon and in vitro and confirms that the panel does not react with non-target organisms. It was tested against more than 38 viral, bacterial, and fungal isolates at high concentration to make sure there is no cross reactivity. And additionally, more than 20 potential interfering substances were evaluated to ensure robust performance even in complex sample conditions.
So beyond this laboratory validation, We also evaluated the performance of the bed fire panel in real world conditions, so a large clinical study was conducted using 680 nasopharyngeal samples from symptomatic horses collected across multiple veterinary clinics in France and the UK. And in addition, we have also included 232 archived samples from a specialised lab that were previously confirmed as positive for at least one pathogen. So each sample was tested in parallel with the Ver panel and the PCS cells from the reference laboratories.
The results showed a positive percent agreement of 95% or higher for the most pathogens EHV1, EHV4, equine flu, and strepy. For equine rhinitis B virus, the agreement was 90%, and for equine adenovirus, 50%. Disease is mainly due to the limited number of positive samples available during this study.
And no result was calculated for equine rhinitis A virus, as no positive samples were found during the study, and we know that this virus is mostly found in North America. In terms of specificity or negative percent agreement, the panel which I'm sorry, 95% or higher for 5 pathogens. The slightly lower values were observed for strepicy and equine rhinitis B.
Likely due to limitations of PCR testing performed by reference laboratories and sample degradation during transport. So overall, this field study confirms that the Vet fire panel delivers reliable and actionable results, even in the dynamic and sometimes challenging conditions of equine care. So it's reinforces the robustness of the system for everyday veterinary practise.
So now I'd like to share an exciting new development that will soon be available for veterinarians. So during this presentation, you understood that we have a very technical technology which includes a full extraction and a very accurate PCR testing. So this enables us to have results regarding the presence of major respiratory pathogens, and this is already a great benefit.
But what else could generate value based on these results? In fact, there is a lot of additional information in this test, and this is why we developed the fireworks suite. So in QW, Fireworks brings all your Vet Fire test data in one secure close-base platform.
And the main interesting feature of fireworks is first the test viewer that gives you access to all your bed fire panel test results. You have the ability to have access to a variety of information provided for each test result, and for positive results, you have also, you also have the ability to view CP value, melt peaks, and melting temperature values. Another interesting feature is the ability by the users to set and receive notifications of specified pathogen detections directly to your inbox or a phone.
And the last but not least exciting feature within fireworks is pathogen surveillance. So this gives users the ability to view and monitor pathogens circulation trends from the test results. And if you choose to share your data, the fireworks web portals gives you access to local, regional, and global trends, so you can easily.
Compare your site's detection rates with what's happening around you, which supports early anomaly detection, timely investigation, and better anticipation of emerging threats. So to, to conclude, The Veg Fire Equine respiratory panel offers a complete, rapid, and easy to use solution, delivering highly accurate results in under 20 minutes, any day, any time, and by anyone. So it's a real game changer in equine respiratory diagnostics.
It's fast, reliable, and ready for the field. Thank you. Hi everybody.
My name is Jane Darcy. I'm here to give a quick overview as an end user of the Ved Fire by Bio Mario. Just a quick summary of myself and the team here at Equiquine Diagnostic Laboratory.
I spent my 1st 13 years at the Irish Equine centre in the meat and bone meal Laboratory analysing animal feed by light microscopy for the presence of animal proteins and fish. I also do testing for animal feed for ruminant DNA by real-time PCR. I have worked in the micro lab in PCR, screening for Clems pneumonia, Tayorella equi genitalis, Pseudomonas sarinosa, strep Equi, and RE coli by real-time PCR.
During my time at the Irish Equiquine centre, I completed a master's by research of antibiotic resistance and vierlin strains of rhotococcus ei in the Irish equine population. I completed an internal auditing course and Lane 6 Sigma. I started my current role at Equiqui Diagnostic Laboratory in 2019, as an analyst.
And during this time, we obviously had COVID, and we carry out a lot of routine testing, including running of bloods, microbiology, PCR, and Eliza. The laboratory is involved in different proficiency testing schemes, including VCA, and the laboratory services, both ambulatory and hospital vets. We have a large range of equids including sport horses, leisure and thoroughbreds.
We currently have a team of 8 staff members with a range of qualifications, including veterinarian, people with equine science, food science, bioscience, and microbiology degrees, who all add a variety of valuable skills to our team here. The laboratory carries out, as I said, a large range of testing, including PCR, LISA, cytology, microbiology, biochemistry, and haematology. The lab, we are constantly looking to explore ways to provide a better service to our vets here.
I suppose, why, why did we choose the R fire? So, this is what we are here to talk about. When we were looking into it, obviously, Bio Mario is a well-known company in operation for many years.
Then the Biofire, which is, kind of, I suppose. The of the vet fire has been used in human hospitals for many years, identifying respiratory pathogens in humans. So this is not new technology.
It's just been applied to veterinary, which is something that we really like here that we know it has been well tested, well run, and, and has good feedback, obviously from the hospitals for many years. As a laboratory, we are always looking at ways to futureproof and keep up with any new diagnostic tests that are available. It is a constantly changing scene with machines and and new.
Ways to analyse with PCRs definitely being one of the ways that is growing exponentially, including different types of technology and lamp and PCR. As a group, we're eager to improve patient outcomes, and having a point of care system on site helps reduce a lot of risk factors. We are constantly monitoring results for any potential disease clusters and this system is really going to help us in in this.
I'm pretty sure Frederick will go into more detail on that. For us, and the speed of execution is, is a, is a critical factor. One of the highlights of the vet fire is, is this, is the speed, it's turnaround of of a result in less than 20 minutes.
And, and that is true. It is, it is very quick to prepare the sample and the running does take less than 20 minutes. Vet fire samples, I suppose prior to this were sent to an external laboratory for testing.
So samples were received into the laboratory, would have been logged into our sample management system and then transported to the external laboratory, possibly overnight arriving the following day. And during this time, the following day, samples will be processed with possibly 7 different amplifications. And this could take several days for results depending on when the sample arrived in that laboratory.
Now with the vet fire, if the animal is on site, we can get a clinical exam done, a blood taken and analysed, nasopharyngeal swab taken and analysed, and all the results together culminating in potentially a diagnosis or some further testing required, or that the animal needs to be, quarantined. So this gives the vet a large amount of information within an hour or two. So this is a huge, huge factor for us.
I suppose, what, what are the main benefits? So, number one, infection control is a huge part of managing an outbreak. Getting a positive or negative result and acting on this information, whether to isolate or release an animal, the quicker the result, the quicker to isolate and to stop the spread of any potential infection.
And, number 2, I suppose, samples coming in after cutoff in laboratories. Obviously we receive samples during the day at varying times of day. When running traditional PCR, samples are run in large batches and extraction takes place with that batch.
The samples are amplified, the PCR is run. So it is not possible often to go back to the start to retract. This is where the vet fire comes in very handy.
It allows as each sample is run individually, you can start a sample anytime you want. There is no waiting or Missing a cutoff and as it's just each individual sample is wrong. And this has been a huge asset where samples come in late, have missed the cutoff.
We can now run them on the PCR and still have a result within the same day. I suppose number 3, time is a critical factor for animals travelling, be it overseas, to shows, new animals moving from farm to farm, or racing. Get a result back on the same day is a huge benefit.
Number 4, it has increased the testing capabilities of our laboratory and it allows us to offer a rapid service for urgent samples that need that quick turnover that you can't get with traditional PCR, especially not with 7 different targets in one day. It's a, is a huge amount for especially smaller laboratories. Number 5, it allows, I suppose the vet to compare the results of the vet fire to their potential current diagnosis of what they feel the animal has within a short space of time.
Thus, it allows them to either confirm or change their potential diagnosis and potentially, maybe look for other tests to confirm or What other line of testing that they may be going down. Number 6 is the vet fire has shown to be extremely sensitive. I'm sure Frederick will be going into this, and going to the limit of detection of each target.
I suppose we're futureproofing the laboratory. The vet fire, I suppose, has a very small footprint. Space is a premium in all laboratories.
So having the module of a small footprint is, of great benefit. And, the touch screen is attached to the module, so it is not taking up more space like often a lot of our instruments would have separate laptops which are taking up more space, but the actual user screen is attached to the module. So that's another space saving issue.
One, the module is, stackable, so you can increase the testing capabilities. Of the lab by stacking a further module on top. I believe it is up to 4, but I'm not 100% sure on that.
What also we like is that the reagents for the test come in a container in a pouch. And in that container is all the PCR reagents and extraction requirements. And these can be stored at room temperature, which reduces the amount of need for fridges and freezers, which most other reagents require and for shipping.
They do not need he. So if they're shipped over the weekend, there is no worry about putting them in freezers and fridges, which is a huge benefit. Which we like, the due to the vet fire extracting and running the PCR only.
Only one room is required in traditional. PCR labs, several separate areas are required to minimise contamination, often with your extraction, your master mix, your PCR, your thermo cycler, so several different areas. As the PCR is contained into the one module, including your extraction and your PCR, you only actually need one area which is of a huge benefit.
And as I said, it's a small, small area that is required. The pouch where the reaction takes place is enclosed, and this also minimises the risk of contamination, so you are actually only loading it once and do not need to touch it once it's being loaded. And, the internal controls are included in the pouch, so this helps to risk, reduce the risk of false, negatives, which we like, and obviously if the internal control fails, the warning will come up telling you that there is an issue that this needs to be repeated.
Influenza is monitored by Bio Mario for any changes to the structure genome. This is something that we really like, so that if there is any changes to it, that they can adjust the primers. In, in, in the, in the mix to take this into consideration.
Yes, that's the kind of main points. And the other thing we like is that the laboratory has the ability to review the data, and not just the end result for in-depth analysis. So you can look at it not just from a, it's positive or negative, a little bit more detail.
The system is extremely user friendly. We have found training people very easy. Obviously, most of the staff here have done traditional PCR, so this is, it's just maintaining .
The cleanliness when dealing with contamination risk. There is only a very small amount of hands-on requirement when loading, and so it's really just involving prepping your nasopharyngeal swabbing, your PBS or VTM buffer, and the colour coding on the reagents in the pouch minimises the risk of putting the sample into the wrong location. So from an ease of use, size.
Sensitivity, specificity, we have found great results with it so far. And obviously we are ongoing running samples through. Yeah, so thank you very much for taking time to listen to me.
Thank you so much, Fleur, Frederick, and Jane, for presenting today's webinar. What a great informative session. We have had a few questions submitted, so we'll get through what we can.
We've got some few minutes remaining. The first question we have is, for Jane. So firstly, how long does it take to train a team, on using the system?
I would say it was very quick. We had Frederick on site. It was, obviously extremely knowledgeable on Bio Mario and the Ved Fire, and so the training is, it is pretty quick, I suppose.
They have made it so user friendly, but as I said, most of our lab staff have already been trained in classical PCR, so it is a little bit easier and I suppose the biggest thing is just maintaining, A good level of cleanliness, decontamination, I suppose, and I suppose is the biggest thing, and that you just don't contaminate from the start, and your disposal technique and gloves, but from a training perspective is very minimal amount of of prep is all I would say of it lovely, and then next question we've got is For Frederick, how many tests can be run in one hour if you have got, maybe a 4 module system? Yes, great question. So, I would like to say it's well organised.
It's, it's possible to run up to 12 pouches per hour using 4 modules. So to note that once, once a pouch is, is loaded, the test run can be initiated within 60 minutes. So this means you can prepare some pouches in advance and change the runs one after another.
Alright, perfect, let me have a look, another one. S And oh another one for you as well, er, what is the difference between this PCR and a lamp PCR? Oh, that is a great, question.
by definition, a PCR, PCR requires a temperature cycling to amplify DNA, whereas a lamp, amplifies DNA at a constant temperature. So it's commonly accepted that lamp is faster and more resistant to inhibitors than PCR. So it may be true for certain one-step PCR methods such as a Tachman PCR, but Vetfire use an ultrafast nested PCR, so that is highly robust against many bishops and delivers results in a very, very short time.
And it also may be important to note that the quality of the sample preparation prior to the amplification step is a key factor in the overall performance of the molecular test. OK. There we go.
Thank you. What other ones have we got here, we. W for Jane, is there much maintenance work that you need to do for the system?
So far for us, we've haven't had no real maintenance on it, and I'm sure Frederick will probably go into more detail on it, but from our side, it doesn't have any maintenance that we've had to do to it because I suppose it's, it has its containment for its speed beaten section and obviously some of the other technology which I would really bond over to Frederick, I'm sure on the thermo cyclers, but . I from when we were talking, there isn't any requirement of us. So I might hand it over to Frederick, you might be able to explain more.
Mm, yeah, the, the, the system is there with, so there are very few, . She, she met us and she Sure, we shouldn't, but that is your question again you know sure. Yeah, well, for Mars we've had no maintenance on it.
We've had no, anything to compare with, but we've had nothing to do to it from its arrival on site and, so far it's so good. So, yeah. And we've got a question for Fleur.
Have you or your team set up or helped set up, an, EIDS type system in any third world countries? We obviously advise internationally on a regular basis if people reach out. I don't know.
Richard's actually in the call as well. He's been with the group an awful lot longer than I have, whether he can on the history the years prior, but not in the time that 78 years I've been with the group have we proactively gone in and conducted surveillance within a country through EIDS oversight, but we, we do advise. Yeah, I'll just yeah, it, it, it's a challenge and we've seen major outbreaks of disease in particularly, parts of sub-Saharan Africa, and the real challenge is getting the diagnostics, getting the diagnosis, so I think these technologies actually offer a great deal of potential to help with that, and as er Frederick has said, I think the, the chances for getting those data.
Out in terms of, connecting to systems like ours are, are there, as long as there are the sort of base, technologies in, in country, to, to do that, and we are aware of other earlier systems used by Welcome, etc. That, that, that can do that. So I think there are, there are, there is greater potential now for setting these systems up and we know that er WA World Organisation for Animal Health, they, they monitor these important diseases as well so it's a it's actually a great question and we would be very happy to speak to either the countries themselves or we also have charitable organisations that do great work in these countries as well and they're obviously keen to get diagnoses very quickly in order that they can.
Seek advice, you know, we've worked with these charities with major flu outbreaks, for example, so yeah, perhaps the, the record in history is not that great, but I think the potential for the future is great. Perfect. Well, well, well, I think that pretty much literally just wraps us up now anyway, but, I just wanna say thank you again, to everyone on, today's webinar.
So thank you, Fleur, Frederick, and Jane. And, thank you again to, Bao and Mario, for sponsoring today's webinar, and, we hope you all enjoyed today's webinar and thank you all for joining us at the webinar there. And er we hope to see you all again soon, so thank you for watching.
Thank you. Take care.
