Molecular analysis of cell-free DNA (cfDNA), small fragments of DNA circulating in blood plasma provide information on the genetic state of normal and tumor cells. Due to its non-invasive nature, molecular interrogation of cfDNA is becoming increasingly popular for tumor characterization and treatment monitoring. The purpose of this study on cfDNA extraction/isolation market is to understand prospective and current market trends at grass root level along with market size, supply, demand information on the end-user application and current business environment. In addition, the study also includes exhaustive information on competitive landscape, restrains and opportunities and other vital information regarding global cfDNA extraction/isolation market.
cfDNA extraction/isolation market by Product
• Isolation Kits
cfDNA extraction/isolation market by Application
• Other Disease
EMA cfDNA extraction/isolation market by Region
• North America
• Asia Pacific
• Latin America
• Cell-Free DNA are fragments of DNA which circulate in the blood fluid and can be utilized as valuable biomarkers. It is a new technology in which small quantity of liquid sample (saliva, urine, or blood) from the patient is collected to examine the freely flowing DNA. Cell-Free DNA (cfDNA) is a noninvasive screening test used to help detect graft rejection, identify mutations in cancer patients and detect various chromosomal abnormalities in the fetus along with the gender of the fetus. The discovery of cfDNA has opened up to new potentials in the field of oncology, gynecology, transplantation and infectious diseases.
• Increased incidences of diseases like chromosomal abnormalities due to increase in maternal age, changing lifestyle, unhealthy diet habits and rising prevalence of cancer along with increasing healthcare expenditure and rising healthcare awareness are the reasons driving the cell-free DNA market. However, there are some other factors such as lack of skilled healthcare professionals in developing markets, high test cost, non-affordability in low income countries, ethical issues related to genetic testing, lack of standardization, unfavorable reimbursement policies and legal and regulatory guidelines are some of the reasons hampering the growth of the market.
• The presence of cell free DNA (cfDNA) in blood plasma was discovered in 1948 by Mandel and Metais. Seventeen years later in 1965, Bendich et al hypothesized that cancer-derived cfDNA could be involved in metastasis. However, it took another year to discover the first link to the disease. In 1966, Tan et al observed high levels of circulating cell-free DNA (cfDNA) in the blood of systemic lupus erythematosus patients. Eleven years later – in 1977 – Leon et al used radioimmunochemistry to demonstrate that for at least half of cancer patients the level of cfDNA in their blood was significantly higher than in normal control subjects.
• The technological progress of the 1990s fuelled by the Human Genome Project allowed more direct demonstration for a tumour origin of at least some cancer patient cfDNA. Arguably the most successful application of cfDNA studies was the discovery of the high admixture of fetal-derived cfDNA in mother’s blood stream by Lo et al. Later, the same group demonstrated that in 70% of women bearing male fetuses fetal Y-chromosome sequences could be detected in just 10 μl of blood plasma. This discovery opened up a new avenue for development of fetal cfDNA-based prenatal genetic testing (NIPT). Recent advancement has opened up new avenue of growth for cfDNA applications in graft rejection, cancer, cardiovascular and autoimmune diseases.
• Multiple properties of cfDNA suggest cell death as its’ major origin. Importantly, cfDNA is double stranded and highly fragmented with most molecules being approximately 150 bp in length . This matches the length of DNA occupied by a nucleosome – the primary unit for spatial organization of DNA in the nucleus. Moreover, the other fragment length peaks correspond well with linear progression of nucleosome units. Interestingly, there is still controversy on whether the higher or lower integrity of cfDNA is associated with cancer. In 2003 Wang et al reported that the comparisons of the relative amounts of 100 and 400 bp PCR products of the β-actin gene demonstrated increased cfDNA integrity in 61 patients with breast and gynecological cancers compared to 65 non-neoplastic patients. This observation is supported by the studies in numerous cancer types summarized in. However, there are also conflicting reports, including by Madhavan et al (1998), who determined that decreased cfDNA integrity (defined as the ratio of concentrations of long, ~260bp Alu and LINE fragments to short, ~100bp, fragments determined by quantitative PCR (qPCR)) correlates with worse outcome. The authors noted that the decreased cfDNA integrity would imply higher apoptotic rates, and that increased apoptosis correlates with higher tumour proliferation. This in turn would imply, that the apoptotic, not necrotic cells are the main source of cfDNA at least in cancer patients.
• Despite almost 70 years of history the biological function of cfDNA has only been unambiguously established for immune response and blood coagulation and its’ function in other conditions, if any, remains nebulous. Neutrophils in human blood release so called neutrophil extracellular traps (NETs) as one of responses to bacterial infection. cfDNA is an important component of these NETs, that allows them to bind and trap microbial pathogens. The release of DNA from neutrophils is thought to occur via an alternative mechanism of cell death termed NETosis. Briefly, two mechanisms are being considered, with the first involving dissolution of the plasma and nuclear membranes that is followed by release of the chromatin into extracellular space. Unlike apoptosis, NETosis does not result in display of the phagocyte activating signals, so the neutrophils that undergo NETosis do not get cleared from the blood stream by phagocytes. An alternative theory postulates the existence of a DNA/serine nuclease extrusion mechanism from intact neutrophils and that autophagy contributes to NETosis. In any case, NETs appear to require intact chromatin lattices. Importantly, cfDNA in NETs triggers blood coagulation, a process that clearly needs to be tightly controlled.
• Currently, although a number of methods for extraction of cfDNA are employed, the efficiency and yield are still low due to loss of starting materials during extraction, and its quantification is variable because of lack of standardization . Nevertheless, for healthy subjects, the average concentration of circulating cfDNA is found to be 10-30 ng/mL with values of cancer patients exceeding 100 ng/mL of plasma [11-13]. The estimation of cfDNA contributed by tumors using multiple methods is between 0.01 to 90%. Most cfDNA fragments measured between 150 to 200 base pairs in length, with a variable half-life in the circulation ranging from 15 minutes to several hours. The amount of cfDNA is influenced by tumor progression, turnover of tumor, tumor size, as well as clearance, degradation, and filtering by the blood and lymphatic circulation. The most common published extraction methods for cfDNA are the commercially available spin column extraction kits. Other reported methods of extraction include magnetic beads, phenol/ chloroform extraction, and alkaline salting. The efficiency of cfDNA extraction can directly impact the outcome of mutation detection i.e., assay sensitivity.