Bordetella Detection & Species Identification Educational Guide
Bordetella Detection & Species Identification Educational Guide
Bordetella Detection and Species Identification with the Vivalytic
Cases of Bordetella infections are rising across Europe. Bordetella species are responsible for whooping cough, or pertussis, which literally means violet cough. Vaccine deployment in the 1940s saw a reduction in the morbidity and mortality associated with these infections and now, healthy adults can be expected to make a full recovery. However, vulnerable populations, such as children, the elderly and the immunocompromised, have been shown to be at increased risk of more severe and long-lasting side effects, including increased risk of mortality.
Traditional methods of identifying Bordetella infections take the form of culture, which can take up to 7 days due to the fastidious and slow-growing nature of these bacteria and provide limited sensitivity1,2. To provide a faster and more sensitive method for the identification of whooping cough pathogens, Randox, in partnership with Bosch, are proud to introduce the Vivalytic Bordetella Cartridge. This real-time PCR assay allows detection of B. pertussis, B. parapertussis and B. holmesii on the Vivalytic system, a universal, fully automated, cartridge-based platform enabling high-plex and low-plex testing, providing an all-in-one solution for molecular diagnostics.
To help you understand the implications of Bordetella infections and those of the Vivalytic system, we have produced a new educational guide, covering the Bordetella species responsible for whooping cough; the pathophysiology and complications associated with these infections; the Vivalytic platform and the benefits it can bring to your laboratory; and finally, a summary of findings presented at ESCMID 2024 in which the Vivalytic Bordetella cartridge showed excellent results. Here, we present this educational guide and a summary of its contents. You can download this guide for free by clicking the download link below.
The Scale of the Bordetella Problem
The rates of positive identification of Bordetella infection are increasing throughout Europe. In England, between January and March 2024, there were 2793 laboratory confirmed cases of whooping cough causing the deaths of 5 infants, compared with a total of 858 cases in 20233. A rudimentary projection model estimates that without intervention, whooping cough cases in the England could total over 15,000 cases by the end of 2024. Rising cases are not isolated to the UK – increased rates of diagnosis have also been reported in Denmark, Spain, and Croatia4. Increased numbers of infections illustrate the need for novel and rapid diagnostics to identify those who have been infected and help reduce the transmission of these bacteria.
Bordetella genus
Bacteria of the Bordetella genus are gram-negative coccobacilli5 which are important pathogens in human medicine as they colonise the respiratory tract leading to a range of pulmonary and bronchial infections6. There are 3 main species associated with whooping cough: of B. pertussis, B. parapertussis (Classical Bordetella) and B. holmesii (pertussis-like disease pathogen).
Pertussis is caused by Classical Bordetella: B. pertussis and B. parapertussis. Despite widespread vaccination cases are rising, partially due to waning immunity. Pertussis is highly contagious and particularly dangerous for infants, who account for most pertussis-related deaths. The disease progresses through three phases: catarrhal (cold-like symptoms), paroxysmal (severe coughing fits), and convalescent (persistent cough). Classical Bordetella species share over 98% DNA sequence similarity and share many crucial virulence factors like toxins adenylate cyclase toxin (ACT), pertussis toxin (PXT), and dermonecrotic toxin5 yet there are variations in potential hosts and disease. For example, B. pertussis is an exclusively human pathogen, whereas B. parapertussis can infect both humans and sheep6.
Bordetella holmesii causes pertussis-like symptoms but is ofen less severe. Unlike classical Bordetella, B. holmesii can cause bacteraemia, especially in immunocompromised individuals. Accurate diagnosis of B. holmesii remains challenging due to its similarities with other Bordetella species.
Whooping cough can lead to complications such as pneumonia, which may develop if fever persists beyond the catarrhal phase2. CNS complications like seizures and encephalopathy occur in less than 2% of cases, often due to hypoxia, hypoglycaemia, toxins, or secondary infections2. Bordetella toxins, especially PXT, increase histamine sensitivity and insulin secretion. Infants are especially at risk of bradycardia, hypotension, and cardiac arrest.
Vivalytic Bordetella Cartridge
To enhance the detection and species identification of Bordetella, Randox introduces the Vivalytic Bordetella cartridge. This user-friendly assay is designed to detect B. pertussis, B. parapertussis, and B. holmesii from a single nasopharyngeal swab or aspirate sample. Utilising Real-time PCR, it enables rapid and accurate detection up to four weeks after symptom onset, differentiating between human pathogenic Bordetella species. With a time to result of just 47 minutes, this assay is invaluable for patient diagnosis and the containment of Bordetella, helping to reduce aerogenic transmission.
Summary of Benefits:
- Sample Volume – 300μl.
- Sample Type – Nasopharyngeal swab sample or aspirates.
- Real-time PCR detection.
- Time to result – ~47 minutes.
- Detection of B. pertussis, B. parapertussis, and B. holmesii.
Rapid and Accurate Detection of Whooping Cough in Clinical Samples
Zimmerman, 2024
At the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) 2024 congress, the Vivalytic Bordetella array showed excellent performance, with a 97.7% concordance and a 97.9% positive percent agreement (PPA) with the reference method.7 It accurately identified all additional positive samples and maintained over 98% PPA across spiked samples, even at low levels. The system’s invalid result rate was notably low at 0.6%, compared to 2.9% with the BioGX assay7.
The conclusions drawn from this investigation are as follows:
- The Vivalytic Bordetella cartridge provided excellent concordance with a sensitive reference method and delivered fast and accurate results.
- This assay is ideal for both hospital laboratories and outpatient settings, thanks to its user-friendly design and quick turnaround times.
- Early identification of infected patients will aid in preventing the spread of re-emerging whooping cough epidemics.
Conclusion
As Bordetella infections rise across Europe, rapid and accurate detection is crucial. The Vivalytic Bordetella Cartridge offers a fast, reliable solution, identifying B. pertussis, B. parapertussis, and B. holmesii with high accuracy in just 47 minutes. This advanced diagnostic tool can help reduce transmission and manage whooping cough effectively.
Take control of your diagnostic capabilities and ensure the best care for your patients. Download our comprehensive educational guide to learn more about Bordetella infections and the benefits of the Vivalytic system.
For more information on the Vivalytic, the panels mentioned, or any of our products, don’t hesitate to reach out to us at marketing@randox.com.
References
- Lauria AM, Zabbo CP. Pertussis. StatPearls Publishing; 2024. Accessed June 12, 2024. https://www.ncbi.nlm.nih.gov/books/NBK519008/
- Pittet LF, Posfay-Barbe KM. Bordetella holmesii: Still Emerging and Elusive 20 Years On. Microbiol Spectr. 2016;4(2). doi:10.1128/microbiolspec.EI10-0003-2015
- UK Health and Security Agency. Confirmed Cases of Pertussis in England by Month.; 2024.
- Smout E, Mellon D, Rae M. Whooping cough rises sharply in UK and Europe. BMJ. Published online April 2, 2024:q736. doi:10.1136/bmj.q736
- Rivera I, Linz B, Harvill ET. Evolution and Conservation of Bordetella Intracellular Survival in Eukaryotic Host Cells. Front Microbiol. 2020;11. doi:10.3389/fmicb.2020.557819
- Hamidou Soumana I, Linz B, Harvill ET. Environmental Origin of the Genus Bordetella. Front Microbiol. 2017;8. doi:10.3389/fmicb.2017.00028
- Zimmerman S. Rapid and Accurate Detection of Whooping Cough in Clinical Samples.; 2024.
Combating Gastroenteritis – Advanced Diagnostic Techniques for Effective Management
Gastroenteritis, often referred to as stomach flu or a stomach bug, affects millions globally each year with symptoms such as diarrhoea, vomiting, abdominal pain, and fever. It is primarily caused by viral and bacterial infections, with rotavirus, norovirus, and Clostridium difficile being the main culprits.
At Randox, we’re dedicated to improving healthcare worldwide. That’s why we’ve produced an educational guide on gastroenteritis and the latest advancements in diagnostic techniques, including a range of novel gastroenteritis test for the Vivalytic POCT system. In this blog, we’ll look at a few of the key points raised in our latest educational guide. You can download this educational guide by clicking the below.
Why Gastroenteritis Matters
Gastroenteritis can lead to severe dehydration, especially in vulnerable groups like children and the elderly. It spreads mainly through the faecal-oral route, which includes consuming contaminated food and water. Prompt and accurate diagnosis is crucial for effective management.
Key Pathogens
Rotavirus
Rotavirus is a major cause of severe gastroenteritis in children. Highly contagious, it leads to rapid dehydration, making rehydration and supportive care essential. Vaccines like Rotarix and RotaTeq are effective in preventing infections.
Norovirus
Norovirus is responsible for most viral gastroenteritis outbreaks. Extremely contagious, it spreads quickly through direct contact and contaminated food. Symptoms include sudden vomiting and diarrhoea, often leading to dehydration. While there’s no specific treatment, staying hydrated is key.
Clostridium difficile
Clostridium difficile, or C. diff, is a leading cause of antibiotic-associated diarrhoea, particularly in healthcare settings. It produces toxins that cause inflammation and damage to the colon, requiring targeted antibiotic treatment for severe cases.
Advanced Diagnostics: The Vivalytic System
Accurate and timely detection of gastroenteritis pathogens is crucial for effective patient management. The Vivalytic Point of Care Testing (POCT) system, developed by Bosch Healthcare Solutions and Randox Laboratories, offers rapid and reliable diagnostics. This system helps healthcare professionals make quicker decisions, improving patient outcomes.
The Vivalytic Gastroenteritis Panels
The Vivalytic panels detailed in our guide include tests for rotavirus, norovirus, and Clostridium difficile. These panels utilise advanced molecular techniques to provide quick and accurate results, helping to streamline the diagnosis process and enhance patient care. By using these panels, healthcare providers can efficiently identify the specific pathogens responsible for gastroenteritis, allowing for targeted treatment and improved patient outcomes.
Features of the Vivalytic System
The Vivalytic system is user-friendly and efficient. It supports both High-Plex and Low-Plex testing, allowing for the simultaneous detection of multiple pathogens from a single sample. This versatility makes it an invaluable tool for healthcare professionals.
Conclusion
Gastroenteritis, caused by pathogens like rotavirus, norovirus, and Clostridium difficile, presents significant health challenges. Advanced diagnostic technologies, such as the Vivalytic system, are crucial in managing and controlling this condition. For a comprehensive understanding of gastroenteritis and innovative diagnostic techniques, download our detailed educational guide.
For more information on the Vivalytic, the panels mentioned, or any of our products, don’t hesitate to reach out to us at marketing@randox.com
Randox’s Sustainable Success with carbonfit
In the dynamic diagnostic testing sector, Randox is leading the charge towards sustainability with a clear vision and strategic partnership with carbonfit. Randox have a commitment to reducing carbon emissions and embracing renewable energy solutions which is setting a new standard for environmental responsibility within the industry.
- Aiming for complete elimination of emissions by 2050 whilst hitting milestones in the years leading up to this.
- Additionally, they are targeting a remarkable 95% reduction in Scope 3 emissions by 2050 whilst hitting milestones in the years leading up to this.
- Additionally, they are targeting a remarkable 95% reduction in Scope 3 emissions by 2050. These goals highlight Randox’s proactive approach to mitigating their environmental impact and aligning with global sustainability objectives.
Partnering with carbonfit, Randox has implemented a comprehensive carbon reduction plan that has yielded significant results. They have identified and implemented energy-saving measures that resulted substantial savings of c.600,000 kWh and a notable reduction of over 1,800 metric tonnes of C02 equivalent (TCO2e). These efforts have not only contributed to environmental sustainability but have also led to financial savings of approximately Ā£180,000 per year, demonstrating the synergies between sustainability and business efficiency.
Randox’s commitment to renewable energy is evident through their planned installation of solar PV systems in 2023 and 2024. By embracing clean energy technologies, Randox is further reducing their carbon footprint and contributing to a more sustainable energy landscape.
One of the main drivers for Randox to work with carbonfit was the need for a more efficient and advanced approach to managing carbon and energy. Transitioning away from manual spreadsheet-based methods, Randox leveraged carbonfit’s sophisticated platform for real-time data management, regulatory requirements including ESOS (Energy Savings Opportunity Scheme), PPN06/21 (Publicly Available Specification for the Assessment of the Life Cycle Greenhouse Gas Emissions of Goods and Services), and SECR (Streamlined Energy and Carbon Reporting), empowering Randox with streamlined operations and enhanced sustainability practices.
Randox’s partnership with carbonfit exemplifies their dedication to innovation, sustainability leadership, and responsible corporate practices. Together, they are driving meaningful change towards a greener future, where environmental stewardship and operational excellence go hand in hand.
PCR rapid tests for Candida auris for Vivalytic by Bosch now available
World’s first fully automated PCR test for detection of the multidrug-resistant fungus at the point of care.Ā
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Vivalytic test detects Candida auris in under an hour at the point of care, making it the world’s first test suitable for screenings.
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Candida aurisĀ infection rates are on the rise in Germany and can cause severe infections, for example in the bloodstream (sepsis).
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Hospitals can use the new screening test to detect colonization, allowing them to implement measures to contain outbreaks.
Bosch Healthcare solutions has developed a PCR test for detecting Candida auris (C. auris)Ā and on the Vivalytic platform. The test is a global innovation now available for order from distribution partners including Randox Laboratories Ltd. and R-Biopharm. This test enables the fully automated detection of the frequently multi-resistant fungus in less than an hour at the point of care. The rapid testing capability also makes it suitable for carrying out screenings when necessary. In contrast, traditional culture tests in centralized laboratories require one to three days, delaying diagnosis and the initiation of targeted treatment. āConsidering the heightened risk of severe progression in individuals with pre-existing conditions, we have developed a new test that enables clinics to respond more swiftly,ā states Marc Meier, managing director of Bosch Healthcare Solutions. Patients with compromised immune systems, such as those in intensive care, individuals with serious underlying conditions such as diabetes, or those who are immunosuppressed due to cancer or HIV, as well as patients about to undergo invasive surgery, face a heightened risk of active infection with C. auris. The mortality rate for C. auris infections ranges from 30 to 72 percent.
āCandida aurisĀ can be transmitted from person to person through contact and contaminated surfaces. When this fungus presents, rapid detection is therefore rapid detection is therefore paramount to enable implementation of effective control and prevention strategies,” says Dr. med. Alexander Maximillian Aldejohann,Ā deputy head of the WĆ¼rzburg Laboratory at the National Reference Center for Invasive Fungal Infections (NRZMyk.) Since July 2023, Germany has implemented a limited statutory reporting requirement under the Infection Protection Act. Aldejohann is in favor of extending this reporting obligation: āThe fungus has the capacity to rapidly develop resistance to many common antifungal agents coupled with the ability to survive for a relatively long time on surfaces. This high so-called tenacity also increases the risk of outbreaks that are difficult to contain.ā
Increasing spread of Candida auris
C. auris is spreading globally. In some states in the U.S, the annual incidence rate has been shown to increase by a factor of 2 to 3. The Robert Koch Institute (RKI) also drew attention to a rise in cases within Germany during the past year in the Epidemiological Bulletin at the at the beginning of May. The RKI points out that in specific areas screening could be beneficial. In the U.S, the annual case count has in the meantime reached the thousands. The Centres for Disease Control and Prevention (CDC) already consider the screening of patients, visitors, and staff for C. auris as a crucial strategy to curb its spread in healthcare settings. While the fungus is harmless for healthy individuals, it can it can lead to severe nosocomial infections, i.e. infections acquired in hospitals or other healthcare environments, in patients at high risk and, if the fungus enters the bloodstream, can trigger sepsis.
Easy handling, rapid detection The Vivalytic Analyser enables effortless testing directly at the point of care: The sample is placed into the test cartridge, which already contains all necessary reagents. The cartridge is then inserted into the Vivalytic Analyser for automated processing. Healthcare professionals require only minimal training to use the system, and the fully automated process significantly lowers the risk of infection. The Vivalytic Analyser thus facilitates rapid and precise diagnostics in PCR quality, bypassing the frequently lengthy process through a central laboratory. Bosch Healthcare Solutions is expecting CE certification for the Vivalytic C. auris test soon.
For More Information Please Contact:
Martin Conway, Phone: +44 (0) 28 9442 2413
E-mail: martin.conway@randox.co
Dementia Action Week 2024
Dementia Action Week 2024 (13th ā 20th May)
The term Dementia describes the different brain disorders that trigger a loss of brain function. These conditions are all usually progressive and eventually severe. Alzheimerās Disease is the most common type of dementia, affecting 62 per cent of all those diagnosed.
Dementia is a general term for loss of memory, language, problem-solving and other thinking abilities that are severe enough to interfere with daily life.Ā Common symptoms include memory loss, confusion, and speech problems. Early warning signs may also include finding it difficult to follow conversations, or programs on TV, forgetting names of friends, or everyday objects and feeling confused even in a familiar environment.
Mainly affecting older people, after the age of 65, the likelihood of developing dementia roughly doubles every five years ā however, for some dementia can develop earlier, presenting different issues for the person affected, their carer and their family. There is also a considerable economic cost associated with the disease estimated at Ā£23 billion a year, which is predicted to triple by 2040. This is more than the cost of cancer, heart disease, and stroke.
At Randox, we recognise the importance in diagnosing dementia early. Ā Through our Randox Ā Alzheimer’s Disease Array which can be used for Rapid Identification of Alzheimerās Disease Risk. Randoxās Alzheimerās Disease Risk Array can be used for the direct determination of ApoE4 status from plasma, eliminating the need for genetic testing, assisting in clinical research and personalised medicine strategies. At Randox, we believe the importance of measuring ApoE4 protein expression in plasma is the way forward to screen those individuals at increased risk of Alzheimer Disease, as new beta amyloid-targeting therapies for this condition are being expected.
Race Against Dementia have been the partner charity for the Randox Grand National Festival over the past two years, working alongside this charity which was founded by three-times Formula One World Champion, Sir Jackie Stewart, with the aim of funding much needed pioneering research into the prevention and cure of dementia.
For further information about the Randox Alzheimerās Array please emailĀ info@randoxbiosciences.com
Serum Indices – Product Spotlight
Errors can occur at any point in the pre-analytical, analytical, or post-analytical stages of a diagnostic test. It is general practice for errors in the analytical stage to be identified through quality control procedures. However, pre-analytical errors are often treated with less importance than those in later stages of testing. Interference caused by haemolysis, icterus and lipemia (HIL) are common forms of pre-analytical error which affect assay methods, yielding erroneous results. The Randox Acusera Serum Indices (SI) control is designed to monitor an IVD instrument’s response in the detection of HIL interferences.
HIL interference is not novel and has been historically identified through a series of visual assessments. While haemolytic, icteric and lipemic interference causes a visual change in the sample, these methods are not quantitative and are subject to interpretation by laboratory professionals. Modern analysers have built-in capabilities for the automated detection of HIL interference which can quantitatively or semi-quantitatively measure haemolysis, icterus and lipemia, and provide and an index for each. This data can then be used to determine if a sample should be accepted for testing or rejected due to intrinsic interference.
The pre-analytical phase of laboratory testing includes collection, handling, transportation, storage, and preparation of samples. Even when the highest level of care is taken to ensure that all aspects of the pre-analytical phase are suitable and correct, errors can occur, exhibiting the need for clear and efficient quality control processes.
As part of our Acusera quality control range, Randox has developed the Serum Indices quality control to aid in the detection of the common pre-analytical error’s haemolysis, icterus and lipemia, collectively known as HIL. HIL interference can have disastrous effects on the quantification of many analytes, and it is therefore vital to determine levels of interference to improve laboratory efficiency and reduce the frequency of erroneous results.
The graph below shows the wavelengths at which each of these interferents may affect assays and the table below describes these forms of interference:
Classical determination of HIL interference took the form of a visual assessment. A sample was examined for tell-tale signs of one or more of these types of interference. However, these methods are subject to operator interpretation and lack harmonisation and uniformity across the industry. These signs are detailed in the table and illustrated in the graphic below:
Modern clinical chemistry analysers have onboard HIL detection capabilities which offer objective, semi-qualitative or qualitative analysis of these forms of interference in a more precise and consistent manner. Automation of HIL detection improves laboratory throughput along with test turnaround times and enhances the reportability of the results.
Errors at any stage of the analytical process will result in retesting of the sample. Errors in the pre-analytical phase can have repercussions such as increased cost of repeated sample collection and testing, poor test turnaround times, and more seriously, delayed or incorrect diagnosis causing an exacerbation in the condition of the patient. To add to the adverse outcomes on patients, repeated testing places additional stress on laboratory resources and staff which ultimately affects every aspect of a laboratory’s daily activities.
To correctly analyse HIL interference, absorbance readings at different strategically selected wavelengths supplement the calculation of the interference indices. C56-A recommends laboratories consider several parameters when selecting an HIL interference analysis method:
Before implementing results obtained from any method detecting HIL in patient samples, it is imperative to evaluate the specificity and sensitivity of the method at a minimum of two clinically relevant concentrations. This assessment should encompass the sensitivity of the icterus index to haemoglobin and lipids, the haemolysis index to bilirubin and lipids, and the lipemic index to haemoglobin and bilirubin.
In instances of HIL interference, laboratories bear the responsibility of managing the associated results and samples. It is crucial never to utilise an HIL index for the correction of patient results. Typically, if a sample is determined to be affected by one or more of these interferences, the laboratory should reject the result and appropriately dispose of the sample. Nonetheless, in certain scenarios, threshold values can be established. For instance, haemolysis may exert a lesser impact on samples with elevated analyte concentrations. In such cases, laboratories may opt for a distinct procedure in handling these results compared to those exhibiting haemolytic interference at lower analyte concentrations.
Acusera Serum Indices Control
The Randox Acusera Serum Indices (SI) control is designed to be used to monitor an IVD instrument’s response in the detection of haemolyzed, icteric and lipemic (HIL) samples. This control can be utilised in laboratory interference testing to assist in improving error detection of pre-analytical errors affecting clinical chemistry testing. This control provides a full range of clinically relevant testing levels, including a negative (-) and three positives (+, ++ & +++).
The Randox Control offers a comprehensive solution with 3 levels for each form of interference and a negative control, providing a wider coverage compared to alternatives in the market. Our product is conveniently supplied in a lyophilized format, ensuring an extended shelf-life and ease of storage. Customers appreciate the stability of our control, as it consistently meets the 14-day open stability claims, minimizing waste and optimizing laboratory efficiency.
Typical Values
RIQAS Serum Indices External Quality Assessment
The RIQAS Serum Indices EQA programme is designed for the pre-analytical assessment of Haemolytic, Icteric and Lipemic (HIL) interferences. Available in a bi-monthly format with the option to report either quantitative or semi-quantitative results for the HIL parameters, this programme also provides an assessment on how these interferences impact on up to 25 routine chemistry parameters. This provides invaluable information on whether a correct judgement is being made to report results.
• Lyophilised for enhanced stability
• Human based serum ensuring commutable sample matrix
• Bi-monthly reporting
• HIL parameters include the option of quantitative or semi-quantitative reporting
• Interpretation of chemistry parameter results
• Submit results and view reports online via RIQAS.net
How can Randox help?
It is crucial laboratories test for haemolysis, icterus and lipemia to ensure the accuracy of their test processes are maintained. ISO 15189:2022 promotes the identification and control of non-conformities in the pre-analytical process, therefore, using Randox Serum Indices control and RIQAS Serum Indices EQA will help laboratories fulfil the requirements of the new edition of this standard.
Randox Serum Indices control displays improved consolidation, stability, and commutability to ensure laboratories are equipped to accurately determine pre-analytical interferences. Our Serum Indices control can be used with most major chemistry analysers including Roche, Abbot, Beckman, Ortho, and Siemens. When used in conjunction with Acusera 24.7, this control offers laboratories the ability to compare their HIL results with their peer group and identify potential failures in their pre-analytical process.
Simply send us an email by clicking the link below and we will get in touch!
Medical Laboratory Professionals Week 2024
Medical Laboratory Professionals Week (MLPW) is recognised every year in the last full week of April. It’s an opportunity to increase the public understanding of, and appreciation for, the hard work of clinical laboratory staff around the world. It’s also an opportunity to inject a little fun into the laboratory. So, this year, we’ve created a Lab Professionals QC Bingo card. Have a go and see how many your laboratory can get!
How many boxes does your lab tick?
If you’re calling Bingo! you must be an Acusera 24.7 customer. If not, keep reading to find out how you can make daily life in your laboratory more straightforward.
What are Medical Laboratory Professionals?
Medicine wouldn’t be where it is today without the work of these laboratory professionals. They’re on the frontline. Around 70% of medical decisions are based on results provided by medical laboratory staff. That’s a lot of pressure on the labs to make sure their results are accurate. Clinical laboratory staff not only perform the tests used to guide diagnosis and disease prevention, but they also check all the tests they use through rigorous quality control (QC) procedures.
This involves testing samples of known values to prove that the test system and its components perform as they should and provide accurate results. To do this, laboratories require QC material. It’s important that what’s in a QC is as similar to what you’d find in a patient sample as possible. This is known as commutability. Good commutability helps limit cross-reactivity in the test and inaccurate results.
It’s also important to make sure the QC material has concentrations of analytes at similar values to those used to make diagnostic decisions. If you wanted to validate the length of the ruler on your desk, it wouldn’t be helpful to set it down on a 100m running track. Similarly, when laboratory professionals want to ensure a test is producing accurate results, they want to test the system at the critical values used to make medical decisions so that they can be confident the results at these values are accurate.
Once lab staff have confirmed the accuracy of their tests, they can begin testing patient samples. For most people, what happens to a sample after it’s taken is a bit of a mystery. MLPW is the perfect opportunity to unravel this a little:
After your sample is collected, it gets sent over to the lab. Even just moving it there needs careful handling to make sure it’s still good for testing when it arrives. Once it’s in the lab, the team checks the equipment to make sure it’s working right and giving accurate results. The QC procedure varies depending on what they’re testing for, but they always make sure their tests are legitimate. Once they’ve checked everything and carried out the tests, a pathologist looks at the results to figure out what’s going on. They use this information to help decide on the best treatment plan for you.
Even this watered-down explanation makes it sound like a lot of work, right? At Randox, we recognise the vital role and dedicated efforts of medical laboratory professionals, and the invaluable contributions they make to society, and we hope that now, you do too.
Acusera 24.7
Bingo! That’s exactly how our customers feel when they realise how much time Acusera 24.7 can save them. Our innovative and intuitive QC data software is cloud-based, allowing you to log in from anywhere in the world to review your QC data.
Along with a wide range of interactive charts, including Levey-Jennings charts, Acusera 24.7 determines measurement uncertainty and sigma metrics for you, saving you the time and stress of manually calculating these tricky statistical analyses. And that’s just the beginning. Acusera 24.7 can link to LIMS for automated data entry, meaning lab staff don’t have to manual type long datasets, unless they want to of course; we also provide both semi-automated data upload and manual data entry options.
Access to a range of reports has never been easier. Acusera 24.7 is particularly useful when gaining or renewing your accreditation, and live peer group QC data, to give additional confidence in the accuracy of your results.
But this article is supposed to be about laboratory professionals, so we won’t bang on about it anymore. We just want everyone to know about Acusera 24.7 so they can get that daily bingo! feeling for themselves. If you want to learn more about our reports, charts, advanced statistical analysis, Acusera 24.7 more generally, or how Acusera 24.7 can help you achieve your accreditation, you can follow the links to the relevant blog post.
Last year, we interviewed two of our laboratory staff, Dean and Meadhbh, to find out what a normal day looked like for them. To find out what a day in the life of a laboratory professional is like, take a look at the interviews here
If you’d like to get in touch with us to discuss the advantages of Acusera 24.7, or you’ve made up your mind and want to get in on the action, reach out to us at marketing@randox.com. We’re always happy to brag about how great Acusera 24.7 is, and how we make life simpler for more and more laboratories every day.
Kate Middleton’s diagnosis reminds us that early detection is key
Kate Middleton’s recent cancer diagnosis has highlighted the importance of early detection to us all. Detecting potential health issues early means individuals can work with healthcare professionals to implement treatment plans, lifestyle changes, or preventative measures. These could mitigate, or even prevent the progression of certain diseases.
An article published by the Daily Mail, which focuses on early cancer detection MOT’s, emphasises the importance of preventative testing and treatment throughout. Charlotte Griffiths, who underwent our Everywoman Plus Tumour-Associated Markers test, stated that it is vital to keep a watchful eye for any unexpected bodily changes, such as unexplained bumps or weight loss, which may indicate a presence of cancer.
At Randox Health, we test for a range of different tumour markers, as well as genetic testing, to deliver information on cancer risk to an individual. This aims to empower people to be proactive towards improving their health and makes them aware of any potential future health risks they may have.
It is estimated that 30-50% of all cancer cases are avoidable, and in many cases, early detection is key and can save lives. Cancer that’s diagnosed at an early stage, when it isn’t too large and hasn’t spread, is more likely to be treated successfully.
Spotting cancer at an early stage can save lives, however many people do not have access to the information or services to facilitate this detection. Over 8 million people in the UK are currently awaiting diagnostic testing. At Randox Health, with over 40 years’ experience in working towards improving healthcare worldwide, we have a focus on preventative healthcare; aiming to achieve better health outcomes whilst reducing the burden on clinical services.
At Randox Health we offer genetic risk tests for inherited cancers, including Breast and Ovarian Cancer, Bowel Cancer, and others. In addition, we offer a range of full-body health tests that include common tumour-associated markers.
The results from your tests allow you to get a more accurate representation of your current health status, as well as giving you the ability to make lifestyle changes to potentially prevent future health risks.
Discover our health checks at randoxhealth.com
RIQAS Performance Assessment – Z Score vs SDI
Z Score vs SDI
You work hard to implement top class quality control in all areas of your laboratory. The success of your labours is reported to you through your External Quality Assessment (EQA) results. It can be frustrating when your report is returned, only for you to find that you’ve been assigned a poor performance score due to other laboratories in your participation group.
At RIQAS, we want your EQA results to reflect your performance, not that of everyone else, to truly illustrate the efficacy of your quality control procedures. This is why, instead of Z scores, we report your performance in terms of a Standard Deviation Index (SDI). However, we know that in some countries, you’re required to report a Z score. Don’t fret. You can still find this result in the .csv file provided with your report.
A Z score is a statistical measurement that describes a value’s relationship to the mean of a group of values. In other words, it’s a value calculated to tell us how many standard deviations (SDs) a result is from the expected mean. Z score is reported in terms of SD’s, therefore a Z score of 0 shows the result is identical to the mean.
While useful in many cases, when used in EQA, a Z score can give a false perception of performance. We want RIQAS participant performance assessment to be based on their individual performance, rather than being impacted by how well, or poorly, the other laboratories in the group performed for a sample.
Z score is calculated using a variable SD. This means that as results are added, the mean and SD can change. For example, if overall performance for a sample improves, the CV associated with the data will decrease, causing an increase in Z score. Let’s take a quick look at how RIQAS performance assessment works, and then we can get into SDI.
RIQAS Performance Assessment.
Our target scoring system has been developed to provide a simple interpretation of your laboratory’s performance. To calculate a target score, your result is calculated as a percentage deviation (V) from the Mean for Comparison. This deviation is then compared to a Target Deviation for Performance Assessment (TDPA) to calculate the Target Score.
The difference between your result and the mean for comparison is expressed as a Target Score (TS) using the following mathematical formulae:
The better your percentage deviation compared to the TDPA, the higher your Target Score will be.
TDPA are set to encourage participants to achieve and maintain acceptable performance. Target Deviations are assigned to be fit-for-purpose and take all possible sources of variation into account, including sample homogeneity and stability as per ISO/IEC17043, ISO13528 and IUPAC.
In general, the TDPA is set so that ~10% laboratories achieve Target Scores less than 50. However, depending on homogeneity and stability, the TDPAs may be adjusted, so that participants’ performance is not adversely affected by sample variability. If your % deviation (V) is equal to the Target Deviation for Performance Assessment (TDPA) then a target score of 50 is achieved.
RIQAS reviews TDPAs annually and the methods used to assign them have been agreed by the RIQAS Advisory Panel.
Standard Deviation Index (SDI)
To provide a more accurate assessment of performance, we use SDI instead of Z score. SDI is a score which compares the participant’s difference from the assigned value (mean for comparison) with an evaluation interval called the Standard Deviation for Performance Assessment (SDPA).
The SDPA calculation involves a series of steps. First, we calculate a CV for Performance assessment (CVPA) as shown below:
As mentioned, the TPDA is normally set so that ~10% of laboratories achieve a TS less than 50. In such cases, the t-value used to convert TDPA to CVPA is ~1.645. However, depending on homogeneity and stability, the TDPA may need be increased, so that participants’ performance is not adversely affected by sample variability. In such cases less than 10% of laboratories will have poor performance, and a larger t-value will be chosen to convert TDPA to CVPA
We then convert CVPA to SDPA:
Using this equation, an initial SDPA is calculated for every mean for comparison (i.e. for all methods, method, and instrument statistics). However, for new parameters or those which have small participation numbers, it’s not always possible to assign a target deviation, TDPA or SDPA. In such cases, the SDPA will be the SD calculated when the mean for comparisons is generated.
According to ISO/IEC17043, when the assigned value is based on consensus (mean for comparison), the uncertainty of the assigned value must be calculated and combined with the SDPA when it is considered to be significant. This forms an adjusted SDPA, which is used to calculate the participant’s performance in terms of SDI.
Using the SDPAadjusted we can calculate SDI using the formula below:
On your RIQAS report, you’ll find the SDI associated with the current sample in the text section of each report page. We also provide your last 20 SDIs, plotted on a Levey-Jennings chart, along with an indication of the mean for comparison for each sample (I = Instrument group, M = Method group, or A = All Methods group). Acceptable performance is an SDI of less than ± 2.
RIQAS EQA
RIQAS is the world’s largest EQA scheme with more than 75,000 laboratory participants spanning over 138 countries. Choosing an EQA provider is no easy task. That’s why we’ve produce a guide to help you find the right one for you. You can download it here.
At RIQAS, we’re always coming up with new ways to make your performance assessment and result interpretation even easier. We’re also proud of our new programmes and pilot schemes. This year, we’re running pilot programmes for Anti-psychotic drugs, Chagas and Blood Typing.
If you’d like to find out more about the range of programmes we provide, visit our website or download our brochure. Alternatively, you can get in touch with us at marketing@randox.com.
Lp(a) Awareness Day 2024
Novel and classical insights into Lp(a) concentration and the effects on various cardiovascular conditions.
Despite advances in understanding and technology, cardiovascular diseases (CVDs) remain a major source of mortality across the world. The World Health Organisation (WHO) estimate that 17.9 million people died due to CVDs in 2019, accounting for around 32% of deaths that year1. First described in 1963, Lipoprotein(a) (Lp(a)) is a macromolecular lipoprotein complex2 which is thought to display proatherogenic, proinflammatory3 and prothrombotic4 potential and is considered an independent causal risk factor for various types of CVD5. These properties provide several mechanisms in which elevated Lp(a) levels may contribute to CVD however the true nature of Lp(a)s relationship to CVD remains largely enigmatic.
Lp(a) concentrations in plasma are principally regulated by variation in LPA gene and levels remain relatively stable throughout one’s lifetime with lifestyle factors having little effect on their concentration6. Due to the highly heritable nature of Lp(a) concentration, those with a family history of Familial Hypercholesterolaemia (FH), elevated LDL-C levels, or Atherosclerotic cardiovascular disease (ASCVD) should be screened, their plasma Lp(a) concentration determined, and their risk of CVD established.
In the last 10 years, there have been many advances in the understanding of this ambiguous lipoprotein which support the causal association with CVD, clarify the established evidence and introduce novel mechanisms of action in relation to Lp(a), shedding light on its obscure pathophysiology. However, there are still diagnostic complications associated with Lp(a) measurement as there is little standardisation in methods of determination5.
Physiology and Genetics
Synthesised mainly in the liver, Lp(a), like LDL, is composed of a lipid centre made of cholesteryl esters and triacylglycerols, surrounded by a shell of phospholipids, free cholesterol, and an apoB-100 molecule. The major difference between other LDL molecules and Lp(a) is the presence of a polymorphic glycoprotein, apo(a), bound to apoB-100 by a single disulphide bond5. It is this apo(a) molecule which contributes to Lp(a)s pathophysiology.
Apo(a) is thought to have evolved from the plasminogen gene (PLG) around 40 million years ago and shares 78-100% sequence homology within the untranslated and coding regions of the fibrinolytic enzyme2. Like plasminogen, apo(a) contains unique domains named kringles5. While plasminogen contains 5 different kringle structures (KI to KV), apo(a) has lost KI to KIII and instead contains several forms of KIV, namely, 1 copy of KIV1 and KIV3-10, 1-40 copies of KIV2, 1 copy of KV and an inactive protein domain at the carboxyl terminus of the molecule7. These hydrophilic subunits are highly polymorphic due to the variation in KIV2 repeats. Individuals may possess two different isoforms of apo(a) one of which will have been passed down from each parent and are expressed codominantly2. These isoforms are dependent on the number of KIV2 repeats they contain2. Isoforms with less KIV2 repeats produce smaller apo(a) isoforms which are found at a higher concentration compared with larger isoforms8 due to the increased rate at which the smaller molecules can be synthesised5. The polymorphisms in KIV2 repeats account for up to 70% of the variation seen in concentration between individuals, with the remainder being attributed to differences in protein folding, transport, and single nucleotide polymorphisms (SNPs)5. SNPs are central in the heterogeneity of apo(a), effecting RNA splicing, nonsense mutations and 5’ untranslated region of the LPA gene resulting in shorter gene translation5,8.
Lp(a) Pathophysiology
Lp(a) is thought to contribute to the risk of CVD through multiple mechanisms. Firstly, Lp(a) molecules display all the same atherosclerotic risk as LDL-C molecules due to their similar fundamental composition, for example, their propensity for oxidisation upon entering the vessel wall, and promotion of atherosclerosis through inflammatory and immunogenic mechanisms 9. However, Lp(a) displays more proatherogenic potential due to the presence of the apo(a) molecule. The structure of apo(a) results in decreased fibrinolysis. Due to its structural similarities, apo(a) competes with plasminogen for binding sites, competitively inhibiting plasminogen, ultimately resulting in reduced fibrinolysis9.
Lp(a) is thought to be a preferential carrier of oxidised phospholipids2 (OxPLs) which covalently bind to apo(a), increase expression of inflammatory proteins, and stimulate the secretion of IL-8 and monocyte chemoattractant protein-1, enhancing its ability to cross the vessel wall9. Some claims require further investigation, however, studies have been carried out which show inhibition of plasminogen activation in the presence of Lp(a)10. It is this indirect mechanism that Lp(a) is thought to conduct its prothrombotic activity8,9.
Clinical Evidence
Many studies have been carried out to determine the association of Lp(a) concentration and CVD risk. Studies such as the Copenhagen General Population Study, the Copenhagen City Heart Study, Dallas Heart Study, and Ischemic Heart Disease Studies provide strong evidence for Lp(a) as a causal risk factor for CVD. Data analysis of the Copenhagen General Population Study reveal that 20% of subjects displayed Lp(a) concentrations of more than 42mg/dl, or around 105nmol/L11, which is considered to result in increased risk of atherosclerotic disease5. It is important to note, there is no accepted conversion factor for converting Lp(a) concentration from mg/dl to nmol/L due to the variability of apo(a) kringles. The unitage will depend on the assay method used5. Another study in a healthcare organisation in Israel showed that Myocardial Infarction (MI) and Coronary Artery Disease was 2.5 times more common in those with high levels of Lp(a) than in the age and sex matched control group3. This study3, along with others5,6,12 describes a linear relationship between Lp(a) concentration and CVD risk, showing at least a 3-fold increase in ASCVD and MI events in adults with Lp(a) concentrations in the top 1% when compared with those in the with concentrations in the bottom 20%3.
The major variation in Lp(a) concentration seen throughout the population, is further evident between ethnicities and sexes. On average, Caucasian subjects display the lowest Lp(a) concentrations, with Black subjects displaying the highest concentrations5. However, the large number of functional variants are consistent across ethnicities suggesting that it is the KIV2 repeats and SNPs that are the major factors contributing to Lp(a) concentration regardless of ethnicity. Lp(a) concentrations are higher in women than men8 with levels increasing post-menopause thought to be caused by a decrease in oestrogen3.
Lp(a) Testing and Screening
The European Atherosclerosis Society (EAS) recommend that all adults are tested at least once in their lifetime to identify individuals who have high levels of Lp(a) and therefore high CVD risk. Screening is also recommended in children who have a family history of Ischaemic stroke, premature ASCVD or high Lp(a) levels in the absence of other identifiable risk factors8. Testing has been associated with reduced mortality rates. This is thought to be because of increased and intensified therapy for those who are identified as high risk due to high plasma Lp(a) concentration6.
There are various assays available for the determination of Lp(a) concentration which vary in accuracy and precision. Many of these assays utilise polyclonal antibodies which recognise different antigenic determinants8. Due to the variability in apo(a) structure and KIV repeats, these assays often overestimate the concentration of large isoforms and underestimate concentration of small isoforms when determining the true Lp(a) levels9. This variation can be partially nullified by using a calibrator series and by selecting a method which is traceable to WHO/IFCC reference material. This allows laboratories to confidently identify individuals considered high risk but may still prove problematic when patients’ results report closer to the assay thresholds.
One study13 compared 5 commercially available Lp(a) assays on an automated clinical chemistry analyser. The assays tested were manufactured by Diazyme, Kamiya, MedTest, Roche, and Randox. The authors show that all the assays tested met the manufacturers claims for sensitivity, linearity, and precision. However, significant bias was observed in 4 out of 5 assays. The only assay which did not display significant bias was the Randox Lp(a) Assay which is traceable to WHO/IFCC reference material. This report highlights the importance of measuring and reporting Lp(a) in molar concentration rather than in mass units to facilitate standardisation and harmonisation in Lp(a) testing13.
Current and Emerging Therapies
Statins are one of the most potent treatments for the primary prevention of ASCVD through the reduction of LDL-C concentration. However, recent studies reveal that statins have no effect on Lp(a) concentration3 and others suggest that statin administration can increase Lp(a) concentration by up to 11%5,9. Nonetheless, EAS do not recommend statin therapy be halted as their strong ameliorative effects on CVD risk are well established and surmount the risk related to increased Lp(a) concentration8.
Niacin (Nicotinic acid) is another established treatment for the reduction of CVD events and act by increasing HDL levels. Niacin can reduce Lp(a) concentration though the reduction of gene expression in a dose-dependent manner5. However, Niacin therapy has not been proven to have beneficial effects on CVD risk8.
A recent metanalysis showed a 26% reduction in serum Lp(a) concentration through treatment with PCSK9 inhibitors. This is thought to be due to a shortage of apoB-100 molecules either because of reduced synthesis or competitive binding with other LDL receptors, resulting in reduced Lp(a) concentration5. Several studies show the efficacy of PCSK9 inhibitors in reducing CVD risk, but this is not yet an approved therapy5,8.
New therapeutic strategies aim to target hepatocytes, the site of apo(a) synthesis, to reduce Lp(a) concentration. Antisense Oligonucleotides (ASOs) inhibit apo(a) mRNA in the nucleus and cytoplasm, ultimately inhibiting Lp(a) secretion5 through the cleavage of the sense strand by ribonuclease H19. While still in clinical trials, ASO therapies show promise in the battle to reduce CVD risk with some studies displaying an overall reduction in Lp(a) concentration of more than 80%9.
Conclusions
There have been major advances in the understanding of Lp(a) pathophysiology in the last 10 years establishing this macromolecular complex as an independent causal risk factor for various forms of CVD, however, more investigation is required to fully understand the mechanisms responsible for this association. With many national healthcare organisations and the EAS recommending universal testing for Lp(a) in adults, more emphasis should be placed on raising awareness of the importance of Lp(a) screening. Finally, more research is needed into therapies which succeed at lowering Lp(a) concentration. While some therapies are in clinical trials, there are currently no approved therapies that achieve this goal.
The Randox Lp(a) assay is calibrated in nmol/L, traceable to the WHO/IFCC reference material, and displays an excellent correlation coefficient of r=0.995 with when compared with other commercially available methods. To accompany this liquid ready-to-use reagent we also offer a dedicated 5 point calibrator with accuracy-based assigned target values (in nmol/l) is available, accurately reflecting the heterogeneity of the apo(a) isoforms.
For more information on this revolutionary assay, visit randox.com/lipoproteina/ or reach out to us at marketing@randox.com.
References
- World Health Organization. Cardiovascular Diseases. World Health Organization. Published June 11, 2021. https://www.who.int/news-room/fact-sheets/detail/cardiovascular-diseases-(cvds)
- Schmidt K, Noureen A, Kronenberg F, Utermann G. Structure, function, and genetics of lipoprotein (a). Journal of Lipid Research. 2016;57(8):1339-1359. doi:https://doi.org/10.1194/jlr.r067314
- Zafrir B, Aker A, Saliba W. Extreme lipoprotein(a) in clinical practice: A cross sectional study. International Journal of Cardiology Cardiovascular Risk and Prevention. 2023;16:200173. doi:https://doi.org/10.1016/j.ijcrp.2023.200173
- Pino BD, Gorini F, Gaggini M, Landi P, Pingitore A, Vassalle C. Lipoprotein(a), Cardiovascular Events and Sex Differences: A Single Cardiological Unit Experience. Journal of Clinical Medicine. 2023;12(3):764. doi:https://doi.org/10.3390/jcm12030764
- Stürzebecher PE, Schorr JJ, Klebs SHG, Laufs U. Trends and consequences of lipoprotein(a) testing: Cross-sectional and longitudinal health insurance claims database analyses. Atherosclerosis. 2023;367:24-33. doi:https://doi.org/10.1016/j.atherosclerosis.2023.01.014
- Lampsas S, Xenou M, Oikonomou E, et al. Lipoprotein(a) in Atherosclerotic Diseases: From Pathophysiology to Diagnosis and Treatment. Molecules. 2023;28(3):969. doi:https://doi.org/10.3390/molecules28030969
- Vuorio A, Watts GF, Schneider WJ, Tsimikas S, Kovanen PT. Familial hypercholesterolemia and elevated lipoprotein(a): double heritable risk and new therapeutic opportunities. Journal of Internal Medicine. 2019;287(1):2-18. doi:https://doi.org/10.1111/joim.12981
- Kronenberg F, Mora S, Stroes ESG, et al. Lipoprotein(a) in atherosclerotic cardiovascular disease and aortic stenosis: a European Atherosclerosis Society consensus statement. European Heart Journal. 2022;43(39):3925-3946. doi:https://doi.org/10.1093/eurheartj/ehac361
- Tsimikas S. A Test in Context: Lipoprotein(a): Diagnosis, Prognosis, Controversies, and Emerging Therapies. Journal of the American College of Cardiology. 2017;69(6):692-711. doi:https://doi.org/10.1016/j.jacc.2016.11.042
- Boffa MB, Koschinsky ML. Lipoprotein (a): truly a direct prothrombotic factor in cardiovascular disease? Journal of Lipid Research. 2016;57(5):745-757. doi:https://doi.org/10.1194/jlr.r060582
- Enkhmaa B, Anuurad E, Berglund L. Lipoprotein (a): impact by ethnicity and environmental and medical conditions. Journal of Lipid Research. 2016;57(7):1111-1125. doi:https://doi.org/10.1194/jlr.r051904
- Svilaas T, Klemsdal TO, Bogsrud MP, et al. High levels of lipoprotein(a) – assessment and treatment. Tidsskrift for Den norske legeforening. Published online January 12, 2023. doi:https://doi.org/10.4045/tidsskr.21.0800
- Wyness SP, Genzen JR. Performance evaluation of five lipoprotein(a) immunoassays on the Roche cobas c501 chemistry analyzer. Practical Laboratory Medicine. 2021;25:e00218. doi:https://doi.org/10.1016/j.plabm.2021.e00218