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.

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 6^th most common in men and 17^th 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.

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