According to the report, global demand for the Digital PCR (dPCR) and Real-time PCR (qPCR) market was valued over USD 4,113.10 Million in 2019 and is expected to reach a CAGR of 8.10% between 2019 and 2025.
The global Digital PCR (dPCR) and Real-time PCR (qPCR) markets are expected to be bolstered by ongoing technological advances in PCR instruments and the rising prevalence of target genetic abnormalities and infectious diseases throughout the projected period. Furthermore, rising grants, investments, and funds will aid the market’s growth in the future years. The growing use of biomarker profiling for disease diagnosis and the successful completion of the Human Genome Project are likely to propel the Digital PCR (dPCR) and Real-time PCR (qPCR) markets forward. The demand for novel Digital PCR (dPCR) and Real-time PCR (qPCR) techniques is expected to increase as R&D activities progress. The shown efficacy of dPCR and qPCR analysis in the estimation and detection of disease-causing microorganisms would boost the use of clinical diagnostic tests like qPCR and dPCR analysis, boosting market growth. The high costs of instruments, particularly dPCR, as well as the technical constraints of PCR, are expected to impede market expansion. Furthermore, the market growth of Digital PCR (dPCR) and Real-time PCR (qPCR) will be hampered by large capital investments and complex scientific validation on a nanoscale level during the development of qPCR and dPCR instruments, as well as the demand for specific reagents depending on the end-use.
Both quantitative PCR (qPCR) and digital PCR (dPCR) can detect and quantify nucleic acids with high sensitivity and precision. Both technologies are similar, but they differ in ways that make one or the other the better fit for certain applications. Learn how qPCR and dPCR stack up in different types of studies.
What are Digital PCR (dPCR) and Real-time PCR (qPCR)?
Real-time PCR, also known as quantitative PCR, is a well-established technique for quantifying nucleic acid in a variety of biological materials quickly and accurately. qPCR is a technique that measures the amount of DNA that accumulates during a PCR reaction. The change in intensity of a fluorescent signal is used to measure the increase in DNA quantity at each cycle. The number of original copies of template DNA in the reaction is determined by comparing it to a reference sample.
Digital PCR is a highly precise method for detecting and quantifying sensitive nucleic acids. For Droplet DigitalTM PCR technique, each sample is divided into thousands of separate reactions (droplets). After end-point PCR cycling, each partition is examined for the presence or absence of a fluorescent signal, and the total number of molecules in the sample is computed. For quantification, dPCR does not require a standard curve. Despite the fact that both qPCR and dPCR enable sensitive detection and precise quantification, their capabilities provide different benefits for different applications. The rapid throughput and wide dynamic range of qPCR make it ideal for screening large quantities of samples. dPCR has unrivaled sensitivity and precision for fractional abundance (mutant/wild-type ratio). Both technologies have complementing capabilities, and combining the two approaches provides researchers with a diverse set of options for genomic applications.
Examine the advantages of dPCR vs. qPCR to see if it’s suited for your research aims.
The main distinction between dPCR and qPCR methods is precision power. While both technologies provide highly sensitive and consistent nucleic acid detection and quantification, Dr. Jim Huggett, Principal Scientist at the National Measurement Laboratory, compares the two technologies to analog versus digital radio. “To acquire the target station with the least amount of interference on an analog radio, the dial must first be fine-tuned.” Still, reception quality is a factor, and the signal is susceptible to static interference. qPCR stands for quantitative polymerase chain reaction. It’s dependable, but it takes some tweaking to achieve a satisfactory result, and even then, there’s background noise to contend with. You simply call up the station on digital radio, and it is either there, with a clear signal, or it is not. The latter is similar to dPCR in that it yields accurate, binary findings. It literally counts the number of DNA molecules present or absent. The great level of precision is due to the clarity of the data mixed with a low mistake rate. Smaller quantitative discrepancies are well-suited to digital PCR.
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- Digital PCR using QIAGEN’s QIAcuity system
Both digital and real-time PCR (qPCR) require a specific time and location for each procedure.
Digital PCR (dPCR) and quantitative real-time PCR (qRT-PCR) are frequently compared by scientists conducting DNA research (qPCR). What’s the best approach to take? Both are correct.
This is the opinion of a Belgian PCR data analysis business, which is increasingly asked this topic. More than a decade after Biogazelle NV sprang out of Ghent University, the Belgian spin-off business still considers quantitative PCR (qPCR) a fundamental technology because of its high throughput and low prices. In response to a growing demand from researchers interested in cancer mutations, copy number variation, and unusual event identification, the company recently purchased a dPCR machine. Biogazelle considers both instruments to be useful reagents. When it comes to reproducibility, scientists rely on qPCR because of the well-established procedures and Minimum Information for the Publication of Quantitative Real-time PCR Experiments (MIQE (1)).
However, digital PCR provides an absolute measurement of the target nucleic acid molecules, as opposed to the relative measurement produced via qPCR. It is possible to measure minuscule variations and correctly quantify minor variants in the background using absolute DNA quantification, which provides for precision, reproducibility, and sensitivity.
With the support of Biogazelle, a biotech company was recently able to develop an experiment for transgene copy number variability using dPCR. Prior qPCR experiments failed to detect fold changes of less than 50%, resulting in unusable data of poor quality. In spite of this, dPCR is capable of picking up subtle changes in copy number, delivering more accurate and dependable results. Both qPCR and dPCR have found a home at Biogazelle. Its researchers choose which system to utilize based on their clients’ needs. In fact, Biogazelle isn’t the only firm that is finding its stride in the PCR industry. Check out what other companies are doing to figure out which PCR technique is best for their needs and how they’re doing it.
qPCR is seen as having a long-term role
Providers are certain that qPCR will continue to be a workhorse method in molecular biology research even as dPCR expands
Why? Researchers have long depended on quantitative polymerase chain reaction (qPCR) because of its speed, sensitivity, specificity and ease of use. Gene expression investigations and validation of other genomic approaches such as DNA microarrays and next generation sequencing (NGS) can benefit from the use of this method for comparison. Researchers may easily access the technology and a substantial corpus of literature for reference because qPCR has been around since 1993.
Researchers can draw from their own historical data when developing and evaluating their research in gene expression analysis, genotyping, pathogen detection, viral quantification, DNA methylation analysis and high resolution melting analysis, among others.
Real-time PCR, as its name suggests, measures PCR amplification in real time. For gene expression analysis, qPCR’s relative nature makes it ideal for determining the concentration or relative abundance of a target relative to a known concentration or control. As with many other biological methods, qPCR data are most useful when compared to other experimental settings, such as sick versus healthy tissue.
Because qPCR assays may detect anywhere from a few copies of a target sequence per reaction up to millions, they provide a wide dynamic range of detection. Because it can detect targets with extremely low and very high copy numbers in the same run, it is ideal for screening and downstream validation tests.
Researchers can choose and select the detection chemical they want to use using qPCR. Intercalating dyes (such as SYBR green) and target-specific probes (such as FITC) can be used in a variety of ways (TaqMan, molecular beacons, etc). Customers can readily alter reaction volume, throughput, and detection method to fit their specific experimental demands, making per-sample cost extremely adaptable.