Internal Quality Control and ISO 15189
Internal Quality Control and ISO 15189
As a major contributor to the IVD industry, like many of you, the trials and tribulations of quality control are an everyday consideration. It is for this reason we strive to make the process of IQC as straightforward as possible. We recognise how busy life in the laboratory can get and believe it is our duty to simplify your QC process as much as possible.
The Acusera range has been designed with this in mind. Our true third-party control range boasts unrivalled levels of consolidation, supplied at clinically relevant concentrations in a suitable, commutable matrix. When used in combination with Acusera 24.7, our interlaboratory management software, the Acusera range will help to reduce analytical errors and maximise precision in your laboratory.
With the recent updates to ISO 15189:2022, we understand that there will be added pressure on many laboratories who are trying to maintain their accreditation. To assist you with your gap analysis and transition to these updated standards, we have produced this accreditation guide, detailing all of the key points relating to this new version of the highly sought after accreditation.
If you’d like to find out more about what we can do to help your laboratory or view our range of Internal Quality Controls, don’t hesitate to contact us at marketing@randox.com or feel free to browse the range on our website https://www.randox.com/laboratory-quality-control-acusera/.
D-3-Hydroxybutyrate & Diabetic Ketoacidosis
Diabetic Ketoacidosis is characterised by an accumulation of ketone bodies in response to insulin deficiency, most commonly occurring in T1DM patients, but is becoming increasingly prevalent among sufferers of T2DM.
Diabetic ketoacidosis is associated with symptoms such as polyuria, polydipsia, fever, vomiting, abdominal pain and fatigue with the most severe cases resulting in disastrous consequences such as cerebral oedema and death.
D-3-Hydroxybutyrate is considered to be the predominant ketone bodies associated with diabetic ketoacidosis and novel methods of detection utilise this biomarker to provide robust and accurate quantification of ketone bodies and aid in confident diagnosis of diabetic ketoacidosis.
This guide discusses the physiological and pathological processes associated with diabetic ketoacidosis and the relevant biomarkers, the complications associated with this condition and classic and novel detection methods.
To download this guide, simply click the image at the top of this post!
For more information on this assay visit https://www.randox.com/d-3-hydroxybutyrate-ranbut/
To read about some of our other superior performance reagents visit https://www.randox.com/superior-performance-and-unique-
Or, to view our wide range of diagnostic solutions visit https://www.randox.com/
Determining bilirubin concentration in paediatric facilities – Vanadate Oxidation Method
The quantification of bilirubin has a wide range of diagnostic utility. In paediatric settings, bilirubin concentrations are commonly used to identify cases of bilirubin encephalopathy or kernicterus.
Historically, bilirubin quantification has been achieved through various techniques derived from the diazo method, first described by Van der Bergh and Muller in 1918. New technologies and novel methods, like the Vanadate Oxidation method, have emerged and have been shown to display superior diagnostic power, driven by its lower sensitivity to interference caused by haemolysis and lipemia when compared with other methods.
This week, we present our educational guide, ‘Determining bilirubin concentrations in paediatric facilities’ which details the key points relating to bilirubin quantification, along with descriptions and comparisons of the methods mentioned above.
To download this guide, simply click the image at the top of this post!
For more information on our Vanadate Oxidation Bilirubin assay visit: www.randox.com/bilirubin
To view our wide range of diagnostic solutions visit: www.randox.com/
Or, if you’d like to discuss this assay, or any of our other products, please contact us at: marketing@randox.com
Identifying and Reducing Pre-analytical Errors in the Medical Laboratory
Medical laboratory professionals must comply with stringent and robust standards in all aspects of their daily activities. The set of standards to which a laboratory must comply will differ depending on the scientific discipline of the laboratory, however, ISO 15189:2022 – Medical Laboratories – Requirements for quality and competence, applies to all medical laboratories. This recent version of the standard introduces increased focus on risk stratification and mitigation for patients and laboratory stakeholders, placing more emphasis on quality control to improve the accuracy and validity of the results obtained.
In a clinical chemistry laboratory, as in others, internal quality control is of upmost importance. Internal quality control (IQC) is the process used to ensure that all results produced are accurate, reliable, and reproducible. To achieve this, a laboratory must carry out checks on the pre-analytical, analytical, and post-analytical phases of testing.
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. Figure 1 shows a graph of wavelengths at which each of these interferents may affect assays and the table below describes these forms of interference:
| Interference | Description |
| Haemolysis | The degradation of red blood cells causes interference between 340-440nm and 540-580nm. Red blood cells experience membrane disruption due to tangential stress which results in degradation of cellular integrity and the release of interfering cellular components such as haemoglobin, K+ ions and aspartate aminotransferase. Haemolytic interference may be evident in assays such iron, lipase, albumin, and creatine kinase. |
| Icterus | Interference as a result of high bilirubin concentrations, affecting assays measured between 400-550nm. The high bilirubin levels result in a yellowish pigmentation of the sample, caused by hepatic necrosis, sepsis, or several other conditions. Most prevalent in neonatal departments, icteric interference can cause inaccuracies in assays for phosphate, creatinine, cholesterol, triglycerides, and uric acid. |
| Lipemia | Interference caused by an aggregation of lipoproteins which affects the turbidity of samples. Lipemic interference can be cause by several mechanisms, the most common being the light scattering effect caused by aggregations of chylomicrons or other large forms of LDL. The larger the LDL molecule, the larger the lipemic effect. Lipemic interference is evident in assays measured between 300-700nm, however, interference increases as wavelength decreases. |
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 below and illustrated in figure 2.
| Interference | Visual indicator |
| Haemolysis | Red discoloration of serum samples which is directly proportional to the concentration of haemoglobin and other interfering erythrocyte components. |
| Icterus | Yellow pigmentation of serum samples increases proportionally to the concentration of conjugated and unconjugated bilirubin. |
| Lipemia | Increased sample turbidity proportional to lipid concentration. |
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.
We hope that by using the Acusera Serum Indices quality control and EQA scheme we can help to improve the accuracy of laboratory testing around the world and remove some of the excessive strain placed on laboratories and the professionals who continually strive for the highest levels of quality in all their work.
How can Randox help?
Randox Sales Reps are experts in their fields and are available to discuss your specific requirements.
Simply send us an email by clicking the link below and we will get in touch!
Medical Laboratory Professionals Week – Industry Insight
As part of our effort to raise awareness of the hard work and dedication displayed by laboratory professionals around the world, we have been talking to individuals from the industry to discover what it is like to work in a medical laboratory environment.
Here, we talk to Dean, a mobile laboratory manager for Randox, based in the UK, to find out what it is like to work in his role.
Q: Let’s start with your name and job title please.
A: I am Dean Gordon and I’m a laboratory manager.
Q: So, what does a normal workday look like for you?
A: A normal day consists of ensuring all our laboratories have everything they need to follow our standard operating procedures and ISO standards. This ranges from ensuring we have enough staff and stock on site to reviewing end of day reports and KPI’s.
Q: What encouraged you to pursue a career in clinical diagnostics?
A: I actually never considered a career in clinical diagnostics. Previously, I worked in marine biology all over the world. During the pandemic I found myself back in Northern Ireland in limbo and Randox were advertising scientific roles on the radio. I thought I would use my science degrees in this moment and work in the lab until the pandemic finished. Over 2 years later, I now find myself still working with Randox and managing ten clinic labs in London and still testing for covid!
Q: What is the most challenging part of your job?
A: The most challenging thing I find is keeping an open line of communication with so many different departments. As our operations have continued to grow over the past 2 years, the more departments you find yourself dealing with, from operations and different clinics to HR and recruitment. There are so many cogs in the wheel and you need to work well with them all to keep it turning!
Q: What is your favourite thing about your role?
A: I love how quickly things move. Since I have started managing labs with Randox, we have opened dozens of new labs and are constantly adding new tests to our portfolio. You always have to be prepared and ready to go when the next new thing is announced. It keeps things exciting. I never feel that I’m bored or standing still in this role.
Q: And finally, why should others consider a career in clinical diagnostics?
A: When you hear feedback from a customer that their test results have helped save or prolong their life and how grateful and happy they are, that they decided to pay for their test – you remember what you are doing can change lives for the better.
We also got the opportunity to speak to Meadhbh, the Randox Clinical Laboratory Services Laboratory Manager, to hear about her work activities and opinions on working in a medical laboratory.
Q: Can you tell us your name and job title please?
My Name is Meadhbh Sheerin, and I am the RCLS Laboratory manager for all of RCLS.
Q: What does a normal workday look like for you at RCLS?
A: Everyday can be slightly different depending on what needs done. But everyday includes morning checks to identify work yet to be completed and ensure target sample turnaround times are met, dealing with customer queries, updating the LIMS system, adding new and bespoke tests to our equipment, managing reagent and other consumables, maintaining up to date SOP and ensuring laboratory staff follow them, and attending in management meetings scheduled. In addition to this I am responsible for hiring and training new staff, setting up new RCLS laboratories and managing the daily activities of other staff.
Q: What encouraged you to pursue a career in clinical diagnostics?
A: For me, it was that people’s health is a priority. Every day, we are saving lives and helping people with their diagnosis, prevent any health conditions, and help them get the right treatment if necessary.
Q: What is the most challenging part of your job?
A: Juggling everything in terms of staff, getting samples in and processed and reports out in time. There is an awful lot to do!
Q: What is your favourite thing about your role?
A: Every day is different and it’s challenging. It is rewarding to know that we are helping individuals to improve their health and that we are the future of diagnostics.
Q: And finally, why should others consider a career in clinical diagnostics?
A: I think everyone should consider a career in some sort of laboratory discipline because you are helping people improve their health and prevent further illness. Preventative care is better than a cure!
Like Dean and Meadhbh, there are millions of conscientious laboratory scientists and technicians which provide crucial testing services all over the world. Working in clinical diagnostics is an incredibly fulfilling career path, providing the opportunity to help people and save lives from a behind-the-scenes yet essential role. We would like to thank Dean and Meadhbh for taking the time out of their busy schedules to answer our questions. Finally, we would like to express our gratitude to all the Medical Laboratory Professionals who have worked tirelessly before, during, and after the pandemic and wish you all the greatest success in the future!
How can Randox help?
Randox Sales Reps are experts in their fields and are available to discuss your specific requirements.
Simply send us an email by clicking the link below and we will get in touch!
Randox Health partners with REVIV
Randox Health have entered into an exciting new partnership with REVIV
Randox Health Clinics are pioneering preventative health care, bringing the world’s most advanced and personalized health programs directly to the public – with the goal of harnessing the power of testing and data to shift healthcare away from sickness management and towards a more proactive approach.
This partnership with REVIV, who championed commercialised IV therapy, will allow people to see real-time results from taking steps to protect their health and to experience the future of wellness.
The IVD drip therapies include:
- The Megaboost, that was designed with wellness in mind this infusion is packed with B vitamins, Vitamin C, Antioxidants and minerals to accomplish restoration of the body’s essential nutrients in one drip.
- The Miniboost, similar to its larger counterpart, the Megaboost, this Miniboost is not to be underestimated! Containing B Vitamins, Vitamin C and antioxidants, it can support energy levels and the immune system whilst aiding protection against cell-damaging free radicals.
- The Royal Flush which supercharges recovery and nutritional balance by providing the ingredients you need directly into your bloodstream. This all-in-one infusion has been directly designed to rehydrate, decreame inflammation and aid detoxification.
- The Hydromax IV which aims to replenish your body’s salts and water.
- The Vitaglow can support detoxification of free radicals that accumulate in the body from exposure to pollutants, daily stresses and chemicals including pesticides.
- The Ultraviv and The Ultraviv pro, both recovery infusions. The Ultraviv can be used to aid recovery against the common cold, sore throats and even the after effects of alcohol. The Ultraviv pro combines a number of prescribed medications with essential vitamins and nutrients delivered alongside the maximum of hydration.
A full list of all REVIV drip therapies and IM booster shots will be available in the Randox Health clinics.
On REVIV partnering with Randox, David Ferguson, Chief Operating Officer, said: “Over the past few years, we’ve seen a dramatic change in people’s behaviours as they seek to understand their health and wellbeing better. At Randox Health, we provide a range of specialised health packages that enable you to take control of your health.
“Our innovative diagnostic technologies can deliver hundreds of results to give you a comprehensive overview of your health and help detect the earliest signs of illness. Collaborating with REVIV is a natural next step, combining our world-class diagnostic services with REVIV signature IV therapies to help our customers protect their current and future health.”
For more information please contact us at: marketing@randox.com
Medical Laboratory Professionals Week 2023
Medical Laboratory Professionals week is taking place from 23rd – 29th April 2023. This is an annual celebration to highlight and acknowledge the contribution of medical laboratory professionals and pathologists to medicine and healthcare. Whether carrying out routine testing or performing vital analysis during states of emergency, patients around the world rely on the hard work and dedication of medical laboratory professionals.
Medical laboratory professionals’ and pathologists’ work often goes unnoticed due to the ‘behind the scenes’ nature of their activities, but today we would like to shine a light on their work and highlight the importance of these individuals to medicine and global health. The role laboratory professionals play in healthcare cannot be understated and Randox would like to give thanks to those around the world who undertake this responsibility every day.
For most people, the process after a sample is taken is largely enigmatic. Therefore, we at Randox would like to elucidate the processes involved and the considerable effort displayed by laboratory staff.
After a sample is taken, it is then transported to a laboratory. Even this supposedly simple process requires careful consideration to ensure the sample is suitable for testing upon reaching the laboratory. Once received, laboratory staff carry out quality control checks to ensure the instrumentation to be used is functioning correctly and providing accurate results. The quality control procedure will differ depending on the scientific discipline but some form of validation of the test process is always required.
Once accurate and robust sample analysis has been carried out a pathologist examines these results or data and works to form a diagnosis. Using this diagnosis, a suitable therapeutic strategy can be determined and administered.
Test results are a major factor in a clinician’s decision for diagnosis and treatment, with 70% of all medical decisions being based on laboratory results. This demonstrates why diagnostics are so important and why Randox believes in celebrating those who make it happen.
As a major contributor to the diagnostics and healthcare industry, we are keenly aware of how important and hard-working medical laboratory professionals are, and the value they bring to the world. This week you’ll find articles featuring a short interview with a medical laboratory professional and a short educational piece on pre-analytical errors.
We hope everyone shares our enthusiasm for celebrating medical laboratory professionals and would like to thank all those who work tirelessly in medical laboratories around the world.
How can Randox help?
Randox Sales Reps are experts in their fields and are available to discuss your specific requirements.
Simply send us an email by clicking the link below and we will get in touch!
The Importance of Maintaining Regular Dietary Patterns to reduce CVD risk
Cardiovascular disease (CVD) is the leading cause of mortality worldwide. An estimated 17.9 million people died from some form of CVD in 2019, accounting for 32% of all-cause mortality that year1. Associations between diet and risk of cardiovascular complications have long been established, largely relating to alterations in lipid profiles.
For as long as anyone can remember, breakfast has been considered the most important meal of the day. Previous studies2 have shown an association between skipping breakfast and increased CVD risk prompting recommendations that up to 30% of one’s daily energy intake should be consumed during the first meal of the day. It has been reported that over 25% of adults skip breakfast. These individuals are often socioeconomically disadvantaged, shift workers, individuals who work particularly long hours, those who suffer from depression or those with poor health literacy2. Another study3 showed that skipping breakfast, when compared with consuming a high-energy breakfast, was associated with a 1.6x and 2.6x higher probability of non-coronary and general atherosclerosis respectively, when all other CVD risk factor had been controlled. This suggests a close relationship between eating breakfast and reducing CVD risk, however, the mechanisms and magnitude of this relationship are poorly understood.
Small, dense low-density lipoprotein cholesterol (sdLDL-C) is a smaller form of LDL-C which boasts greater propensity for uptake by arterial tissue, increased proteoglycan binding, and increased susceptibility for oxidation4. sdLDL-C concentration is strongly associated with CVD risk, yet once again, the mechanisms of this association remain enigmatic. It is thought that all of the metabolic changes associated with alterations in sdLDL-C concentration collectively contribute to the increased risk of CVD, with the main drivers being its propensity for uptake by arterial tissues and its long circulatory stability4
Skipping breakfast and sdLDL-C
A recent study investigated the relationship between skipping breakfast and the effects on lipid parameters5. In a cohort of around 28’000 people from the Japanese population, this study looked at the several markers, including sdLDL-C, to develop an understanding of the importance of regular dietary patterns for reducing the risk of CVD.
The study participants were divided into two main categories: breakfast eaters and breakfast skippers. These categories were further subdivided to differentiate men and women, over and under 55 years old, and those who eat staple products (rice, pasta, bread, etc.) and those who did not. The participants contributed blood samples which were tested for several cardiovascular biomarkers: Creatinine, Liver ALT, Total Cholesterol, Triglycerides, direct LDL-C, HDL-C and sdLDL-C.
They found that around 26% of men and 16.9% of women skipped breakfast regularly. Of these, most were considered young and had significant increases in concentration of triglycerides, LDL-C and sdLDL-C compared with those who ate breakfast almost every day.
Table 1. Median concentration of triglycerides, LDL-C, and sdLDL-C for breakfast skippers and eaters5
| Analyte | Breakfast Skippers (mg/dL) | Breakfast Eaters (mg/dL) |
| Triglycerides | 103 | 93 |
| LDL-C | 124 | 122 |
| sdLDL-C | 34.7 | 32 |
This investigation also revealed that in this cohort, 20% of men and 27.3% of women did not regularly consume staple foods as part of their diet and had higher median sdLDL-C concentration.
Table 2. Median concentration of sdLDL-C in men and women who eat or skip staple food products in their diet5
| Gender | Staple Skippers (mg/dL) | Staple Eaters (mg/dL) |
| Men | 34.1 | 31.6 |
| Women | 25.8 | 24.7 |
The data from this study supports the finding that individuals who skipped breakfast had higher sdLDL-C concentrations than those who ate breakfast consistently. Skipping breakfast can therefore be associated with troublesome lipid parameters in both genders and all age groups in the Japanese population. This study suggests that eating breakfast every day is crucial to maintain beneficial lipid parameters and reduce the risk of developing CVD.
The data also show that individuals who skipped staple foods in their meals presented with higher concentrations of sdLDL-C and a higher sdLDL-C/LDL-C ratio, in men and postmenopausal women, when compared with those who included staple foods in their meals. It is becoming increasingly common to remove staple foods from one’s diet due to their high carbohydrate content and the prevalence of low-carbohydrate diets. This data exhibits the importance of maintaining a nutritionally balanced diet to help reduce the risk of developing CVD.
As the first large scale study of its kind, this analysis provides clear insight into the increased risk of CVD associated with not only skipping breakfast, but failing to maintain a nutritionally balanced diet. The major limitation of this analysis is that it only includes individuals from the Japanese population and the same affects may not be seen in populations from other ethnicities. Therefore, further in-depth analysis is required to confirm these findings in other ethnicities
Randox sdLDL-C Assay
The Randox sdLDL-C assay employs the clearance method which displays good correlation with the gold standard in sdLDL-C quantification, giving laboratories increased confidence in their results first time, every time. Supplied as liquid ready-to-use reagents, this this test can be applied to a wide range of clinical chemistry analysers, producing results in as little as 10 minutes. Relevant controls and calibrators are also available from Randox as part of the Acusera range.
Randox sdLDL-C Assay Key Features
- Direct, automated test for convenience and efficiency.
- Rapid analysis results can be produced in as little as ten minutes, facilitating faster patient diagnosis and treatment plan implementation.
- Liquid ready-to-use reagents for convenience and ease of use.
- Applications available detailing instrument specific settings for a wide range of clinical chemistry analysers.
- sdLDL-C controls and calibrator available.
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)
- Ofori-Asenso R, Owen AJ, Liew D. Skipping Breakfast and the Risk of Cardiovascular Disease and Death: A Systematic Review of Prospective Cohort Studies in Primary Prevention Settings. Journal of Cardiovascular Development and Disease. 2019;6(3):30. doi:https://doi.org/10.3390/jcdd6030030
- Uzhova I, Fuster V, Fernández-Ortiz A, et al. The Importance of Breakfast in Atherosclerosis Disease. Journal of the American College of Cardiology. 2017;70(15):1833-1842. doi:https://doi.org/10.1016/j.jacc.2017.08.027
- Rizvi AA, Stoian AP, Janez A, Rizzo M. Lipoproteins and cardiovascular disease: An update on the clinical significance of atherogenic small, dense LDL and new therapeutical options. Biomedicines. 2021;9:1579. doi:https://doi.org/10.3390/biomedicines9111579
- Arimoto M, Yamamoto Y, Imaoka W, et al. Small dense low-density lipoprotein cholesterol levels in breakfast skippers and staple food skippers. Journal of Atherosclerosis and Thrombosis. 2023;30. doi:https://doi.org/10.5551/jat.64024
For more information on our sdLDL-C assay or any of our other products, please contact us at: marketing@randox.com
World Health Day 2023
Randox is celebrating WORLD HEALTH DAY!
We are dedicated to improving healthcare using innovative diagnostic technologies, for a range of health conditions including heart disease, diabetes, Alzheimer’s disease, cancer, and stroke.
Whilst the science is complex, the applications are not. Diagnostic testing takes place every day behind the scenes of GP surgeries, laboratories, and hospitals.
To celebrate and raise awareness of the health industry, we have written the article below which focuses on the challenges in cancer screening, diagnosis, improving risk stratification, and patient management.
Give it a read and let us know your thoughts!
Overcoming the challenges in cancer screening, diagnosis, improving risk stratification & patient management
The problem
Cancer diagnosis is an art, in many cases requiring complex equipment and time-consuming protocols to achieve only relatively specific and sensitive tests. There are several approaches used to screen for and diagnose different forms of cancer including the identification of biomarkers, quantification of metabolic analytes and genomic sequencing, each displaying their own advantages and limitations.
The identification and quantification of analytes is an effective screening method for some cancers. The Glasgow Prognostic Score (GPS) utilises serum CRP and albumin quantification to provide invaluable prognostic information for pancreatic, colorectal, hepatocellular and other forms of malignant tumours1. While this, and other similar methods can provide reliable, prognostic data they are rarely considered diagnostic. Furthermore, tests such as these often require multiple samples or large sample volumes, repeated hospital visits, and manually dominated test protocols, increasing the risk of human error.
Next generation sequencing (NGS) is an innovative form of genomic sequencing used in cancer diagnosis to identify genes, parts of genes, and genetic mutations known to be related to either cancer in general, or specific forms of cancer. Whilst accurate, NGS screening requires expensive, complex equipment and prolonged protocols, somewhat limiting their utility in providing patients with a timely diagnosis.
Finally, a variety of imaging techniques can be used to visualise tumour growth in the body. These methods are well established, however, are normally not independently diagnostic and can only detect large groups of cancer cells, or tumours, which are evident only in the later, more fatal stages of cancer.
Due to limited resources and other contributing factors, an estimated 1 million cancer diagnosis have been missed in Europe since the beginning of the COVID-19 pandemic2, providing evidence for the need for fast, simple, and accurate screening and diagnostic techniques.
The solution
In 2002, Randox invested £180 million to develop the patented Biochip Array Technology (BAT) in response to the known limitations in diagnostics. This ground-breaking assay technology utilises multiplex testing methodology to provide a rapid, accurate and user-friendly methods for the diagnosis and screening of a wide variety of biomarkers. For use in molecular and protein-based immunoassays, BAT works by combining a panel of related biomarkers in a single biochip with one set of reagents, controls, and calibrators. Unlike other forms of testing which require a sample for each individual test, BAT can provide simultaneous qualitative and quantitative detection of a wide range of biomarkers from a single sample.
The biochip detection system is based on a chemiluminescent reaction. This is the emission of light, without heat, as a result of a chemical reaction. An enzyme is used to catalyse the chemical reaction on the biochip which generates the chemiluminescent signal. The light emitted from the chemiluminescent reaction that takes place in each Discrete Test Regions (DTR) is simultaneously detected and quantified using a Charge-Coupled Device (CCD) Camera.
Each biochip has up to 49 Discrete Test Regions meaning up to 44 tests can be carried out simultaneously. The additional DTRs are reserved for internal quality control and visual reference, a unique Biochip Array Technology feature.
Advantages of Biochip Array Technology
-
Reduced times spent on individual tests as a result of multiplex testing, helping reduce required time and expense .
-
The vast biochip test menu allows clinicians to detect routine and novel markers for advanced diagnostic analysis.
-
Multiple sample types can be used on a single analyser including serum, plasma, whole blood, urine, oral fluid and alternative matrices.
-
Testing for multiple markers helps to simultaneously increase the amount of returned patient information allowing for more informed patient diagnosis.
-
BAT has a proven high standard of accurate test results with CV’s of less than 10%.
-
Barcoded biochips and patient samples ensure complete traceability of results.
-
Biochips are manufactured free from Biotin-streptavidin to reduce cross-reactivity.
Randox BAT has been used to develop several arrays for the detection of routine and novel biomarkers related to various forms of cancer, allowing for improved risk stratification and improve patient management reducing current invasive diagnosis methods.
Randox Pancreatic GlycoMarker Array
Pancreatic cancer is an aggressive form of cancer, one associated with very poor prognosis, often not diagnosed until it has reached the late stages. The 5-year survival rate of 9% attributed to pancreatic cancer indicates a requirement for fast, effective screening and diagnosis. The only FDA approved biomarker for use in pancreatic cancer diagnosis is CA 19-9. However, this biomarker has been shown to display inadequate sensitivity and high levels of false results when used independently and is known to be indicative of various forms of cancer1.
To this end, Randox has developed the Pancreatic GlycoMarker Array, which utilises three distinct biomarkers in a glycosylation-based multiplex detection system. The simultaneous detection of CA 19-9, Carcinoembryonic antigen (CEA) and Alpha-1-Acid Glycoprotein (A1AG) from a single patient sample provides increased sensitivity and specificity for pancreatic cancer when compared with traditional CA 19-9 analysis alone1. Capable of providing results in under 2 hours, this array provides impressive test turnaround times enabling effective intervention and treatment.
| Biomarker | Description |
| CA 19-9 | Cancer antigen 19-9 is a sialyl-Lewis A tetrasaccharide which around 10% of the population cannot express. It is associated with various forms of cancer most importantly, pancreatic, colorectal, and hepatic cancers. Levels of CA 19-9 are also known to be elevated in non-malignant diseases such as chronic pancreatitis1. |
| CEA | Carcinoembryonic antigen is a widely utilised biomarker for different tumours. In pancreatic cancer, increased CEA levels were shown to be evident in 60% of patients3 |
| A1AG | Alpha-1-Acid Glycoprotein is primarily produced by the liver; however, expression has been shown by various cancer cells. Altered glycosylation of A1AG is indicative of malignancy and metastasis4. |
The table below has been taken from an analysis carried out by Randox to determine the Area under curve (AUC), sensitivity, and specificity of these biomarkers, both as a full panel, and individually:
Table 1. Results of an investigation to determine the Area under curve (AUC), sensitivity and specificity of Randox GlycoMarker Array targets both individually and as a panel.
Colorectal Cancer
KRAS, BRAF, PIK3CA Array
Colorectal cancers (CRCs) are the third most common form of cancer, accounting for an estimated 1.93 million cases in 20205. There are three major genes which, when mutations occur, are associated with CRC: KRAS, BRAF and PIK3CA.
Kirsten rat sarcoma (KRAS) is an oncogene frequently mutated in CRC. Around 40% of CRC patients display missense mutations in KRAS most of which occur in codons 12, 13 and 616. The protein encoded by this gene acts as a molecular switch, alternating between a GDP-bound inactive state and a GTP-bound active state. The binding of GTP to the KRAS protein is key in the binding of effectors and the initiation of several downstream pathways which promote cell growth and proliferation. Mutations in the KRAS gene will result in a disruption in hydrolysis of GTP and/or an increase in nucleotide exchange, resulting in an accumulation of the KRAS protein in its active state, the subsequent, continuous activation of downstream signalling pathways and ultimately the proliferation of cancer cells6. Approximately 85% of KRAS mutations occur in codons 12, 13, and 61, with codon 12 being host to 65% of these. Mutations in these codons are associated with extremely poor prognosis compared with wild-type (WT) KRAS cases6.
Mutations in the BRAF gene are evident in an average of 12% of CRC patients, the majority of which are attributed to a BRAF V600E (valine 600 to glutamate) substitution7. CRC patients which display this mutation have a median overall survival (OS) of 11 months and are associated with high levels of epigenetic expression through DNA methylation when compared with WT BRAF patients. V600E mutations are known to inhibit the expression of caudal-type homeobox 2 (CDX2), a tumour suppressor and transcriptional factor crucial in the regulation of intestinal epithelial cell differentiation, cell adhesion, and polarity. The loss of CDX2 activity is associated with high levels of metastasis and poor prognosis in CRC patients7.
PIK3CA mutations are common in various forms of cancer, promoting carcinogenesis through the dysregulation of important cancer signalling pathways. PIK3CA encodes the alpha catalytic subunit of PIK3 (phosphatidylinositol-4,5-bisphosphate 3-kinase), which is responsible for the phosphorylation of phosphatidylinositol-4,5-bisphosphate to phosphatidylinositol-4,5-triphosphate. This newly phosphorylated molecule simultaneously binds kinase PDK1, mTORC2 and serine/threonine kinase, AKT. The phosphorylation of AKT results in the downstream activation of pro-carcinogenic factors and inhibition of tumour suppressor activity, including inhibition of the transcription factor, FOXO1. FOXO1 has several important functions relating to cell apoptosis and proliferation and acts as a context-dependant tumour suppressor8.
The Randox KRAS, BRAF, PIK3CA Array is based on a combination of multiplex PCR and biochip array hybridization for high discrimination between multiple wild‑type and mutant DNA regions in the KRAS, BRAF, and PIK3CA genes. Providing there are enough copies of DNA present, approximately 1% of mutants can be readily detected in a background of wild‑type genomic DNA. A unique primer set is designed for each mutation target and control, which will hybridize to a complementary DTR on the biochip array. Each DTR corresponds to a particular mutation target. With the ability to simultaneously detect 20 mutation points within the KRAS, BRAF and PIK3CA genes, this array can aid clinicians in diagnosis and screening of CRC and help provide insightful information regarding treatment options and prognosis.
Female Bladder Cancer Array
Bladder cancer is considered the most significant cause of haematuria. Bladder cancer is very common, estimated to be the 6th most common in men and 17th most common form of cancer in women9. However, this disparity means bladder cancer in women is often overlooked and the associated haematuria is often attributed to other diagnosis. Those who are correctly diagnosed often experience delayed diagnosis and treatment resulting in worse survival probability10. Cystoscopy, an invasive endoscopy procedure of the urethra and bladder, is the gold standard for the diagnosis. This procedure carries high risk of infection, bleeding and is extremely uncomfortable for the patient. Furthermore, bladder cancer is associated with a high recurrence rate, meaning patients require monitoring for the remainder of their lives, displaying the urgent need for less invasive, fast, effective, and gender-specific screening methods for bladder cancer detection.
The urgent need for evidence-based risk stratification models for screening, diagnosis and subsequent management of patients presenting with haematuria prompted Randox to develop the Female Bladder Cancer Array. Utilising a combination of biomarkers known to provide high sensitivity and specificity, this array is designed to assist clinicians to differentiate patients presenting with haematuria from those with other causes, while removing the need for invasive imaging techniques. This array detects IL-12p70, IL-13, Midkine and Clusterin to provide a comprehensive panel of targets aiding clinicians in risk-stratification, diagnosis, and ongoing monitoring of female bladder cancer patients.
Biomarker |
Description |
IL-12p70 |
Interleukin 12p70 is a disulphide linked heterodimeric cytokine which regulates inflammation by linking innate and adaptive immune responses and potent inducer of antitumor immunity. |
IL-13 |
Interleukin-13 is an immunoregulatory cytokine which plays an important role in carcinogenesis through affecting tumour immunosurveillance. IL-13 in the bladder cancer patients suggests that this cytokine is involved in progression in bladder cancer patients. |
Midkine |
Midkine is a member of a family of heparin-binding growth factors, which has been reported to have an important role in angiogenesis and is associated with bladder cancer progression. |
Clusterin |
Clusterin is conserved glycoprotein that has been distinguished from human fluids and tissues which plays a key role in cellular stress response and survival. It is evident in cancer metastasis, which is particularly important to design the strategies for treating metastatic patients. |
The Evidence Investigator
The Evidence Investigator is a compact semi-automated benchtop analyser. It is a perfect fit for medium throughput laboratories seeking maximum use of bench space without compromising on the volume of samples processed.
-
Estimated turnaround time: Less than 5 hours
-
Detection from nucleic acid
-
Batch testing
-
Suitable for laboratory setting
-
Comprehensive test menu
-
Medium to high throughput – 54 samples and reporting 540 results in less than 5 hours
For references related to this article- References
For more information on this, please contact us at: market@randox.com
MRSA ā Emerging Therapeutic & Screening Approaches
Staphylococcus aureus is a gram positive, commensal bacteria found in normal human flora on the skin and mucous membranes. The commensal nature of this organism results in colonisation of around half of the general population, rising to around 80% in populations of healthcare workers, hospitalised patients and the immunocompromised1. However, given the opportunity to colonise internal tissues or the bloodstream, S. aureus infection can cause serious disease. Skin conditions caused by S. aureus include impetigo, scalded skin syndrome, boils, and abscesses. Examples of more serious conditions include meningitis, pneumonia, endocarditis, bacteraemia, and sepsis2.
Antimicrobial resistance (AMR) has, and continues to be, one of the largest threats to global health. In 2019, it is estimated that 1.27 million deaths globally were directly attributed to AMR, based on the drug-susceptible counterfactual, with only ischaemic heart disease and stroke accounting for more deaths in that year1. Figure 1 shows a global distribution map of MRSA isolates from the data of this comprehensive study. Methicillin-resistant Staphylococcus aureus (MRSA) was first identified only one year after the introduction of the penicillin-like antibiotic, methicillin3. While methicillin is no longer used in clinical practice, the term MRSA is used to encompass resistance to commercially available antibiotics such as β-lactams3. For many years, much work has gone into seeking novel therapies to combat drug-resistant bacteria, however, the indiscriminate overuse of antibiotics seen around the world, along with other factors, continues to contribute to the rise in AMR.
Identification of drug-resistant strains of bacteria is crucial to allow for characterisation of the pathogen and correct treatment of the infection. Classical evaluation consists of a routine culture to verify a diagnosis based on presenting symptoms. However, this can be a time consuming and laborious process which may delay diagnosis and treatment of a potentially fatal infection1.
Methicillin-Resistant Staphylococcus aureus
Methicillin is of a class of antibiotics known as β-lactams which bind to the penicillin binding protein (PBP) of the bacteria. PBP is responsible for crosslinking between N-acetylmuramic acid and N-acetylglucosamine which forms the architecture of the bacterial cell wall. When β-lactams bind to the PBP, a build-up of peptidoglycan precursors triggers autolytic digestion of peptidoglycan, facilitated by hydrolase. This reduction in peptidoglycans results in the loss of the integrity of the bacterial cell wall and ultimately culminates in cell damage caused by high internal osmotic pressure.
While methicillin has lost its clinical utility due to the emergent resistance, MRSA is used to describe S. aureus which displays resistance to penicillin-like antibiotics such as amoxicillin and oxacillin, as well as other forms of commercially available antibiotics like macrolides, tetracyclines, and fluroquinolones4. A meta-analysis by Dadashi et al., showed that 43% of S. aureus isolates where methicillin-resistant, exhibiting the prevalence of MRSA5.
Transmission is possible from direct contact with an infected individual or through contact with fomites2. MRSA infections can be categorised as either community acquired infections (CA-MRSA), or hospital acquired infections (HA-MRSA). While rates of HA-MRSA have fallen over the last ten years, this decrease in infection rates has not translated to CA-MRSA6. This is evidence of the requirement for quicker, easier testing in community settings to identify those infected by MRSA and to trigger the initiation of isolation and treatment.
While the pathophysiology of MRSA will largely depend on the causative strain of bacteria, collectively, S. aureus is the most common bacterial infection in humans and may result in infections of varying severity including1:
- Bacteraemia
- Infective endocarditis
- Skin and soft tissue infections
- Osteomyelitis
- Septic arthritis
- Prosthetic device infections
- Pulmonary infections
- Gastroenteritis
- Meningitis
- Toxic shock syndrome
- UTIs
Development of resistance and resistance mechanisms
Antimicrobial resistance arises from a combination of mechanisms. Genetic mutations are crucial in the development of resistance mechanisms. These genetic mutations must favour the survival of the mutated gene and the advantage of AMR mechanisms to the survival of bacteria cannot be understated. Regarding MRSA, S. aureus can gain resistance through horizontal gene transfer mediated by plasmids, mutations in chromosomal genes or mobile genetic elements4. Methicillin-susceptible Staphylococcus aureus (MSSA) gains the staphylococcal cassette chromosome (SCCmec) gene, a gene containing mecA, which is responsible for some of the resistance mechanisms displayed by MRSA4. The collection of antibiotics the bacteria gains resistance to, will depend on the SCCmec gene type.
The first mechanism of resistance is the expression of β-lactamase which functions to degrade β-lactams, ultimately resulting in loss of function of the antibiotic. This enzyme hydrolyses β-lactam ions in the periplasmic space, denaturing the antibiotic before it can interact with bacteria3. The mecA gene encodes the protein penicillin-binding protein 2a (PBP-2a), a type of PBP which has lower affinity for β-lactams, as well as other penicillin-like antibiotics due its conformation, meaning that the presence of these antimicrobial agents does not confer a loss of structure in the bacterial cell wall1.
One study conducted by Hosseini et al., investigated resistance mechanisms in MRSA and showed that all multidrug resistance MRSA strains displayed biofilm formation as part of its resistance strategy7. Biofilms induce resistance to high concentrations and a large variety of antimicrobial agents and help regulate anti-bacterial immune responses. Biofilm formation is mediated by the protein, polysaccharide intercellular adhesin (PIA). Furthermore, MRSA strains which display biofilm formation are associated with more severe and more virulent infections7.
Current and Emerging Therapeutic Strategies
Other types of antibiotics have been used to treat MRSA infections over the years. Vancomycin has been used to combat infections resistant to penicillin-like antibiotics as they display a different mode of action. Vancomycin inhibits peptidoglycan synthesis by forming hydrogen bonds within the structure of peptidoglycan precursors2. While this strategy has proven effective for past 50 years, more and more strains are displaying vancomycin resistance in addition to resistance to penicillin-like antibiotics8. One study by Deyno et al., estimates the prevalence of vancomycin-resistant S. aureus in Ethiopia to be around 11% 4. Daptomycin is another antibiotic which has been shown to be effective in MRSA treatment. This cyclic lipopeptide binds to the bacterial membrane, resulting in cell death9.
Due to the decreasing number of available, effective antibiotics, novel therapeutic strategies are required to combat MRSA infection. One of the most promising approaches uses antimicrobial peptides (AMPs). AMPs are naturally occurring molecules of the innate immune system and have one of two mechanisms of action: membranolytic action and non-membranolytic action. AMPs normally consist of and amphipathic or cationic structure, between 5-50 amino acids long. Naturally occurring AMPs have been used as a model to develop synthetic AMPs, designed to neutralise the limitations of natural AMPs boasting an improved half-life and improved antimicrobial properties3. Membrane disruptive AMPs can be further categorised by mechanism of action. The first is the Toroidal-pore model in which AMPs form vertical pores in the bacterial membrane causing a change in conformation of the lipid head. Next is the Barrel-stave mode, in which AMPs bind to the bacterial membrane and aggregate before breaching the cell wall causing uncontrolled cell movement, resulting in cell death3. Finally, in the carpet model, the membrane is destroyed in a detergent-like action where the AMPS arrange on the cell membrane with their hydrophobic part facing the phospholipid bilayer, altering the surface tension of the membrane. This eventually results in the formation of micelles and the destruction of the bacterial membrane3.
Non-membrane disruptive AMPs require much more investigation; however, it is accepted that these AMPs enter the cell, reacting with important intracellular components inhibiting protein and nucleic acid synthesis, cell division and protease activity3.
Silver nanoparticles (AgNPs) exhibit broad spectrum antimicrobial properties through various mechanisms of action. These nanosized particles boast increased antimicrobial properties due to an increased surface area per volume ratio. The first mechanism of action to note is AgNPs direct adhesion to the bacterial membrane, which alters the structural integrity of the membrane, allowing the AgNPs to penetrate the cell, wreaking havoc on the intracellular components until it loses the ability to carry out essential cellular processes3.
Once the AgNPs aggregate on the bacterial surface, the difference in electrostatic charge, driven by the positive charge displayed by the AgNPs and negatively charged bacteria, pit formation occurs on the cell surface, inhibiting vital cellular movement, resulting in cell death3. AgNPs may also inhibit protein synthesis by denaturing ribosomes and directly interacting with DNA. This interaction can cause denaturing of the DNA helix and ultimately result in cell death3. Finally, AgNPs can induce the production of reactive oxygen species (ROS) and free radicals. The molecules cause irreversible cell damage to the bacteria3.
While AMPs and AgNPs each possess individual limitations such as toxicity and instability, studies show that a combination of these therapeutic strategies can overcome these issues, stabilising the antimicrobial agents to their respective target sites3.
Screening, Testing & Evaluation
Classical determination of MRSA and other bacterial infections consists of obtaining a patient sample and growing colonies from the patient sample in culture. These cultures can then be investigated under a microscope and characterised, allowing diagnosis and the initiation of treatment. Whilst effective, these methods are time consuming and laborious, taking up to three days for cultures to develop, somewhat limiting their utility for the diagnosis of potentially fatal infections.
New molecular rapid PCR microbiology techniques aid in the identification of bacterial strains through a three-step process involving extraction, amplification, and detection. These new methods allow for timely identification of infectious strains and AMR characterisation. Specific genes or sections of gene which are responsible for AMR can be detected, helping to achieve strain characterisation and aid physicians in prescribing the correct treatment plan. These methods improve test turnaround times to around one to two days and help to reduce the risk of costly human error and contamination.
Vivalytic MRSA/SA
Bosch Vivalytic MRSA/SA is an automated qualitative in vitro diagnostic test based on real-time PCR for the detection and differentiation of methicillin-resistant Staphylococcus aureus (MRSA) and Methicillin-sensitive Staphylococcus aureus (MSSA) DNA from human nasal- or oropharyngeal swabs to aid in the diagnosis of MRSA infection of symptomatic or asymptomatic individuals, providing results in less than 1 hour.
Without MRSA screening, many MRSA colonised patients remain unnoticed in hospitals and will not be isolated. Without Isolation many of these patients transfer the pathogen to at least one other patient during their hospital admission. PCR based screening is associated with high precision and fast time to results and is often used for early decisions on isolation and hygiene measures.
This POCT system provides fast, accurate characterisation of MRSA/SA strains while minimising the required user steps and reducing the need for expensive laboratory equipment helping physicians implement timely and effective treatments.
Detectable Pathogens:
- Methicillin-resistant Staphylococcus aureus
- Methicillin-sensitive Staphylococcus aureus
Specific Gene Targets:
- SCCmec/orfX junction
- MecA/MecC
- SA422
Some of the other benefits of this test include:
- Multiple sample types – Data shows that for approx. 13% of MRSA carriers, the pathogen is only located in the throat. Therefore, using throat swabs significantly increases the sensitivity of detection by approx. 26%.
- Broad MRSA Range – mecA or mecC are the genes responsible for resistance to β-lactam antibiotics. mecA/meC is part of the mobile genetic element Staphylococcal cassette chromosome mec (SCCmec). Vivalytic MRSA/SA can detect mecA as well as mecC and a broad variety of SCCmec elements which help to reduce false negative results.
- Fast time-to-result – Provides quick results in less than 1hr allowing quick decisions on therapies. Traditional culture time-to-result is 48-72hrs and laboratory PCR is 12-24hrs.
- This highly automated system minimises the user steps required to achieve a result while limiting the requirement for expensive lab equipment and sample transportation. Vivalytic MRSA/SA POCT test allow the implementation of treatment as soon as 1hr after sample collection.
References
- Murray CJ, Ikuta KS, Sharara F, et al. Global Burden of Bacterial Antimicrobial Resistance in 2019: A Systematic Analysis. The Lancet. 2022;399(10325):629-655. doi:https://doi.org/10.1016/S0140-6736(21)02724-0
- Nandhini P, Kumar P, Mickymaray S, Alothaim AS, Somasundaram J, Rajan M. Recent Developments in Methicillin-Resistant Staphylococcus aureus (MRSA) Treatment: A Review. Antibiotics. 2022;11(5):606. doi:https://doi.org/10.3390/antibiotics11050606
- Masimen MAA, Harun NA, Maulidiani M, Ismail WIW. Overcoming Methicillin-Resistance Staphylococcus aureus (MRSA) Using Antimicrobial Peptides-Silver Nanoparticles. Antibiotics. 2022;11(7):951. doi:https://doi.org/10.3390/antibiotics11070951
- Liu WT, Chen EZ, Yang L, et al. Emerging resistance mechanisms for 4 types of common anti-MRSA antibiotics in Staphylococcus aureus: A comprehensive review. Microbial Pathogenesis. 2021;156:104915. doi:https://doi.org/10.1016/j.micpath.2021.104915
- Dadashi M, Nasiri MJ, Fallah F, et al. Methicillin-resistant Staphylococcus aureus (MRSA) in Iran: A systematic review and meta-analysis. Journal of Global Antimicrobial Resistance. 2018;12:96-103. doi:https://doi.org/10.1016/j.jgar.2017.09.006
- Kourtis AP, Hatfield K, Baggs J, et al. Vital Signs: Epidemiology and Recent Trends in Methicillin-Resistant and in Methicillin-Susceptible Staphylococcus aureus Bloodstream Infections — United States. MMWR Morbidity and Mortality Weekly Report. 2019;68(9):214-219. doi:https://doi.org/10.15585/mmwr.mm6809e1
- Hosseini M, Shapouri Moghaddam A, Derakhshan S, et al. Correlation Between Biofilm Formation and Antibiotic Resistance in MRSA and MSSA Isolated from Clinical Samples in Iran: A Systematic Review and Meta-Analysis. Microbial Drug Resistance. Published online March 10, 2020. doi:https://doi.org/10.1089/mdr.2020.0001
- Verma R, Verma SK, Rakesh KP, et al. Pyrazole-based analogs as potential antibacterial agents against methicillin-resistance staphylococcus aureus (MRSA) and its SAR elucidation. European Journal of Medicinal Chemistry. 2021;212:113134. doi:https://doi.org/10.1016/j.ejmech.2020.113134
- Deyno S, Fekadu S, Astatkie A. Resistance of Staphylococcus aureus to antimicrobial agents in Ethiopia: a meta-analysis. Antimicrobial Resistance & Infection Control. 2017;6(1). doi:https://doi.org/10.1186/s13756-017-0243-7
