A promising tech­nique for detection of SARS-CoV-2

Instead of being utilized solely for research, less complex tools that rely on cloud computing and AI could change the market.

The Covid-19 pandemic underscores a need to understand the biological mechanisms underlying disease progression and severity. Early studies reported a correlation between immune response dysfunction and disease severity in Covid-19 patients. A deeper understanding of how SARS-CoV-2 interacts with the host immune system is required to better understand Covid-19 pathogenesis and identify biomarkers that scientists could use to predict, monitor, and prevent Covid-19 progression. Flow cytometry, which is routinely used to assess immunophenotypic and functional changes in response to viral infection, plays a critical role in Covid-19 research due to its ability to simultaneously measure the expression of multiple proteins at the single-cell level. Various research teams are using flow cytometry to detect SARS-CoV-2, identifying how the virus interacts with host cells to understand how the immune system responds to the infection.

Recently, December 2021, Biomedical scientists at LMU have found a new marker in the blood of Covid-19 patients. It furnishes insights into the course and development of the disease and could lead to better diagnoses. In the Journal of Extracellular Vesicles, Brocker and colleagues have now reported about the role of phosphatidylserine in Covid-19. A molecule normally found in cell walls, it could be significant for pathophysiological mechanisms relating to the immune system and blood coagulation. And potentially, it could also be suitable as a new biomarker for predicting the severity of the disease by means of a blood test.

All immune cells were analyzed using the phosphatidyl­serine test and separated by means of flow cytometry. The instrument created microscopic images of each cell simultaneously. On the basis of the image files, the researchers were able to recognize whether phosphatidylserine was present – and where it was located. This revealed that the immune cells did not carry the signal inside them.

The measurements also revealed a connection between the severity of Covid-19 and phosphatidylserine. Elevated values during the active phase of Covid-19 correlated strongly with the severity of the disease and could ultimately lead to better diagnoses. As a marker, phosphatidylserine outperformed established lab markers for inflammatory processes in the body, for leukocytes, and for coagulation factors that are currently used for the clinical evaluation of Covid-19. Various laboratory parameters are presently used for classification and form the basis of the WHO scale from zero points (healthy) to eight points (dead from Covid-19).

Brocker’s system is still designed for research laboratories, as very few hospitals have flow cytometers with imaging capabilities.

Therefore, the LMU researchers now want to determine whether ordinary flow cytometers – of the kind that many hospitals have in their laboratories – are also suitable for measurement.

The Indian industry is keen that in a similar manner, flow cytometric evaluation of the immune system on Covid-19 patients be done in India too.

Recently, Metropolis Healthcare announced the launch of a simple blood test that is able to identify the risk status of a hospitalized Covid-19 patient. CoviRisk – TBNK (IVD approved flow cytometry-based lymphocyte subset assay, that estimates T, B, and Natural Killer (NK) cells besides the CD4 and CD8 subpopulations of T-cells in peripheral blood) for cell-mediated response to SARS CoV2 test can assist clinicians to identify severity and prognosis, in triaging of hospitalized Covid-19 patients, and helps to determine if a patient might require IMV (intubation with mechanical ventilation) and ICU facility. Multiple studies and evidence suggest that flow cytometry data can be used to explore the role of T-Cell subtypes as a surrogate biomarker to risk, disease severity, and clinical outcomes for patients suffering from Covid-19. CoviRisk TBNK by flow cytometry can be one of such biomarkers.

Lupin has announced the launch of its diagnostics business in India as part of diversification. The company plans to open over 100 labs and 1000 collection centers across India in the next three years. The company would not just be offering routine diagnostic tests, but also specialty ones, such as a range of tests like flow cytometry, molecular diagnostics, cytogenetics, microbiology, serology, histopathology, immunology, and hematology, among others.

Lupin is not the first pharmaceutical company to enter diagnostics, Mankind Pharma, another large drug maker, has been into diagnostics business with focus on small towns.

Apollo Diagnostics has opened a global reference lab in Hyderabad that will offer over 3000 tests and service clients from across India and Southeast Asia as well. The facility is equipped with a modern molecular biology lab with RT PCRs, CB NAAT, and GenXpert testing and will offer end-to-end services with flow cytometry for routine testing and immunoflourometry.

Aster Labs has launched a pathology lab in Mangaluru. The lab is spread across 1500 sq ft and offers more than 2500 tests ranging from routine to specialized tests. Aster Labs will offer more advanced tests on flow cytometry.

The global flow cytometry market size is expected to reach a value of USD 11.5 billion by 2027, according to Grand View Research, Inc. The market is expected to expand at a CAGR of 8.9 percent from 2021 to 2027. The high prevalence of chronic diseases, the introduction of technologically advanced flow cytometry solutions, and increasing R&D investments in the pharmaceutical industry are expected to propel market growth in coming years.

In 2021, the instruments segment dominated the market for flow cytometry and accounted for the largest revenue share of 35.4 percent, and is expected to maintain its dominance over the next 6 years. This high share is attributed to new advancements in technology and the introduction of novel cytometers by key players. The high price of these instruments is also contributing toward the high revenue generation in this segment. The reagents and consumables segment also held a significant revenue share in 2021 in the market for flow cytometry owing to their increased adoption in diagnostics and research.

The cell-based segment dominated the market for flow cytometry and held the largest revenue share of 76.6 percent in 2021. Increasing demand for early diagnosis and growing awareness pertaining to associated benefits of cell-based assays are factors contributing to its highest share. Furthermore, advancements in technologies of cell-based assays, such as innovation in instruments, labels, affinity reagents, software, and algorithms, are anticipated to drive adoption in the coming years.

The research segment dominated the market and held the largest market share of 49.6 percent in 2021, and is anticipated to hold its dominance over the next six years. This high share can be attributed to the increasing R&D activities pertaining to cancer and infectious diseases, including Covid-19. In addition, the increasing R&D investments in the pharmaceutical and biotechnology industry are also expected to create a conducive environment for market growth.

Technological advancements, resulting in enhanced accuracy, portability, and cost-effectiveness, are expected to present this market with future growth opportunities. Small-sized high-throughput cytometers are anticipated to gain popularity in the near future due to their associated benefits, such as ease of use and cost-effectiveness. Furthermore, improvements in fluorescent dyes and the introduction of bench-top cytometers are factors expected to drive market growth. Rapid advancements in multicolor flow cytometry, which has extensive applications in new drug development, have led to an easy cellular analysis by simultaneous evaluation of several parameters. These devices are extensively adopted by many contract research organizations.

The restraining factors that challenge the growth of the flow cytometry market are the availability of alternate and cheaper techniques, high costs associated with flow cytometry reagents and instruments, budget restrictions for research purposes, and expensive installation and maintenance costs. High costs and the scarcity of labor and skilled technicians in the lab and clinics also hamper the flow cytometry market growth. Additionally, the lack of awareness among the end-users and less accessibility of technical expertise is further restraining the market growth.

The flow cytometry market in the Asia-Pacific region is likely to grow at the highest CAGR, which is greatly due to increasing development and production of new biologics, vaccines, and drugs, expanding medical treatment for infectious and chronic diseases, rising median age of the population, and increasing focus on attracting international and domestic players in the market.

Noteworthy companies operating the global flow cytometry market are Becton, Dickinson, and Company, Beckman Coulter, Inc., Thermo Fisher Scientific, Inc., Merck KGaA, Sysmex Partec GmbH, Luminex Corporation, Miltenyi Biotec GmbH, Bio-Rad Laboratories, Inc., Sony Biotechnology, Life Technologies Corporation Inc., EMD Millipore Corporation, Miltenyi Biotec, Agilent Technologies, Inc. and Affymetix Inc.

The key players operating in the market are adopting various strategies, such as the launch of new products, collaborations, geographical expansion, and partnerships to sustain their position in the market. For instance, in November 2021, Merck announced the launch of its new ColorWheel flow cytometry portfolio, utilizing a proprietary technology optimized for flow cytometry that allows users to independently select antibodies and dyes for assembly in any desired combination.

In November 2021, Bio-Rad has launched ten StarBright Dyes, including StarBright Blue 700, StarBright UltraViolet 400, and eight StarBright Violet Dyes, with many more in the pipeline. These dyes provide multiple benefits for flow cytometry.

In September 2021, Sysmex Europe GmbH has launched its sample-preparation system PS-10 for IVD use in the EMEA region. By thoughtfully automating the sample-preparation work in clinical flow cytometry laboratories, the PS-10 helps establish more efficient workflows – it relieves staff from manual work, eases their burden of documentation, and aids in overall standardizing clinical flow cytometry testing.
In October 2021, BD Life Sciences-Biosciences, a segment of BD (Becton, Dickinson and Company), in collaboration with Christian Medical College, Vellore, has launched its second Center of Excellence (CoE) in flow cytometry for clinical research in India. In July 2021, the company has also launched bdbiosciences.com, an updated digital marketplace for flow cytometry. The new website provides an enhanced online purchasing experience for flow cytometry users and their procurement teams.

In June 2021, Thermo Fisher Scientific announced the launch of the Invitrogen Attune CytPix flow cytometer, an imaging-enhanced flow cytometer that combines acoustic focusing flow cytometry technology with a high-speed camera.

In March 2020, Sony introduced its new fluorescent dyes KIRAVIA Dyes, which are utilized for reagents for many life science applications. These are used for tissue and cells in this field, like immunology and regenerative medicine. These KIRAVIA Dyes are organic polymers that have fluorescent dyes loaded on novel organic polymer developed by Sony.

In January 2020, Miltenyi Biotech introduced the world’s first operator with its new MACS GMP Tyto cartridge, which creates fluorescence-based, GMP-cell sorting compliant opportunities. It is used first on human trials, particularly in people with Parkinson’s disease.

In June 2019, Beckman Coulter Life Sciences had announced that it had acquired Cytobank. This Cytobank offers a software-based platform for multi-parametric single-cell data analysis, and this technology couples with Beckman’s CytoFLEX LX-21-color flow in meeting high complex workflow and requirement of clinical researchers.

In January 2019, Luminex Corporation announced the acquisition of Millipore Sigma’s flow cytometry. This acquisition allows for increasing the flow-based detection systems as well as widening interactions based on microcapillary technologies.

The latest developments in flow cytometry are allowing ever more refined gating of target subpopulations, and thousands of parameters can now potentially be measured per cell to generate unique fingerprints. However, the current approach in advanced flow cytometry is typically to capture all the data first, and then manually (and subjectively) select only a few, specific features for further analysis. Thus, much information of interest may be lost, preventing powerful comparisons of cell subpopulations or disease phenotypes that may never otherwise be visible to the researcher. Therefore, there is a pressing need for objective, automated, and robust data analysis tools, and, to gain widespread use, user-friendly software interfaces. Arguably, for this reason, advanced flow cytometry remains primarily a research, rather than a clinical, tool.

However, to overcome these drawbacks, flow cytometry is being combined with other technologies to get the best of both, in one equipment.

The biggest game changer will be the use of artificial intelligence (AI) in flow cytometry. AI seems to be the ideal solution to save time, labor, and give objective results. FCM captures vast amount of data but when analyzed manually, only a few features are selected for further analysis, losing other useful data. AI can give meaning to the vast data acquired by FCM.

Image flowcytometry (IFC) combines the imaging data of immune-fluorescence with the high-throughput quantitative data of FCM and helps in localization of the marker within the cell. Introduction of deep neural networks in AI can revolutionize IFC. The large number of single-cell images, obtained through IFC, can be used to train the networks to rapidly identify cells, and diagnose common disorders/identify rare cells and bring it out from research to clinical diagnostics.

Imaging cell-sorter flow cytometry can accelerate drug discovery in pharmaceutical field as only events of interest are collected (instead of getting lost in the vast data) to be analyzed further.

Mass cytometry combines FCM with mass spectrometry. It uses elemental metal isotopes, conjugated to monoclonal antibodies, to evaluate around 50 parameters simultaneously on individual cells with minimal overlap between channels.

Combination of flow cytometry with immunomagnetic separation detects microbial contaminants in food samples. This may be extended to clinical samples to give rapid results overcoming the shortcomings of traditional cultures.

Spectral analyzers are the next generation of multicolor flow cytometers, with unparalleled sensitivity and detection range. These are extremely useful for separating labels with narrow-emission spectra.

Flowcytometry-on-a-chip would be the dream come true owing to its low manufacturing cost, drastic reduction in the size of the flow cytometer, and bringing flowcytometry to the bedside for point-of-care diagnosis.

With rapid amalgamation of various other technologies, flow cytometry has the potential to be the best clinical tool for diagnosis and monitoring of diseases.

The present improvements in medical research, as well as those in flow cytometric technology’s diagnostic capacities, will cause a shift in the flow cytometric business landscape. Instead of being utilized solely for research, less complex tools that rely on cloud computing and AI to assist medical workers in making diagnostic and therapeutic decisions for patients could change the market.