Identifying mutations present in a small subset of DNA templates is critical to advances in several areas of biomedical research. For example, rare mutation detection can be used to help identify residual disease at surgical margins or in lymph nodes, to track treatment progress when assessed in plasma, and to identify individuals with early surgically curable disease when assessed in stool, sputum, plasma, and other body fluids. patient. Numerous studies have shown that identifying rare mutations is critical for optimizing treatment.
Molecular cloning and sequencing provide a new dimension to rare mutation analysis, as they allow the identification of the type of mutation, not just its presence. One of the powerful new methods is based on digital PCR (dPCR), which works by dividing a DNA or cDNA sample into many separate, parallel PCR reactions, some of which contain the target molecule (positive) and others that do not. (feminine). A single molecule can be amplified a million-fold or more. During amplification, TaqMan chemistries and dye-labeled probes can be used to detect targets of specific sequences.
Screening of lung cancer patient plasma for EGFR L858R, L861Q, Del19 and T790M mutations using a dPCR dual assay.
Our dPCR is an accurate, cost-effective and well-established method that overcomes common limitations of qPCR, such as the need for a standard curve, low accuracy when measuring rare targets, and lack of sensitivity under high background conditions. Several embodiments of our dPCR include analysis of molecules arranged in multiwell plates, polony, microfluidic devices and water-in-oil emulsions. These techniques are used to accurately identify mutated templates through binding to oligonucleotides specific to potentially mutated bases.
Creative Biogene uses dPCR to rapidly detect and quantify rare mutations in low-occurrence targets in cancer research samples, reliably identifying significant cancer-related mutations. For liquid biopsy analysis, treatment response and resistance can be monitored by detecting relevant cancer driver and resistance mutations.
With its simple and reliable features, our digital PCR has shown strong advantages in the fields of cancer molecular marker discovery, infectious disease research, genome structure variation analysis, gene expression analysis and other fields. If you would like to learn more about rare mutation analysis, please contact us.