Quantifying Genomic Variation

Digital PCR with Higher Accuracy and Consistency Enables Breakthroughs in Quantitation and Monitoring

Natural variability of the human genome can range from single nucleotide polymorphisms (SNPs) to insertions or deletions of entire regions that lead to copy number variations (CNVs). Since CNVs affect large regions of the genome and can include part of a gene or even multiple genes, their impact on human health has been widely studied.
Digital PCR, which utilizes simple counting of target molecules to achieve absolute quantification, enables higher precision and accuracy to detect and quantify copy number variations – even if the changes are small. The Combinati Absolute Q provides four optical channels for multiplexing, enabling more genes to be interrogated per reaction – requiring fewer samples when compared to qPCR. Early identification of copy number profiles that are related to genetic conditions can facilitate timely interventions that could lead to better patient outcomes.

Quantifying Severity of Spinal Muscular Atrophy

Spinal muscular atrophy (SMA) is the leading genetic cause of infant mortality. The disease is largely caused by the loss of the SMN1 gene. A duplicate gene – SMN2 – has been found to produce partial function. The presence of SMN2 can partially compensate for the loss of SMN1 and reduce disease severity.
Leveraging digital PCR’s unparalleled accuracy in quantifying copy number variation (CNV), Combinati developed the first 4-plex dPCR assay to measure SMN1, SMN2 and total SMN using the RPPH1 gene as reference. The assay was validated using 15 clinical samples including a 5cp SMN2 sample.

Precision cancer monitoring

For applications in translational cancer research, the Absolute Q digital PCR platform brings 4-color multiplexing capabilities, improved sensitivity, and a one-step, walkaway workflow to your research. This publication (Scientific Reports), demonstrates the Absolute Q’s capability to detect and quantify non-small cell lung carcinoma (NSCLC) rare genetic mutants (EGFR T790M) with precision cell-free DNA (cfDNA) standards.
Additionally, this publication highlights how the Absolute Q was used to validate the complete molecular remission of a patient diagnosed with juvenile myelomonocytic leukemia (JMML) by tracking a tumor specific (CML) fusion gene (BCR-ABL1) with a custom assay. The assay which was validated down to 0.01% mutant allele frequency was used to precisely track the quantity of the cancer mutation over time.

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