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Digital PCR Detection of EGFR Mutations in cfDNA from Clinical Samples

Digital PCR Detection of Mutant Alleles -- Tech Note
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This study evaluated the detection of EGFR L858R and T790M cancer mutations in 4 mL non-small cell lung cancer (NSCLC) diagnostic remnant plasma samples collected in K2EDTA tubes, deidentified and acquired from Discovery Life Sciences and Precision for Medicine. The NEAT Liquid Biopsy Kit was used for sample extraction on the KingFisher™ Apex System, followed by analysis on the QIAGEN QIAcuity Digital PCR System with LNA mutation assays. These samples were identified as EGFR mutation-positive by examining treatment data gathered at the time of blood collection. Samples 1, 3, and 4 were expected to be positive for EGFR T790M with other potential EGFR mutations, and Sample 2 was expected to be positive for EGFR L858R but not T790M.

The data showed that all four samples characterized as true positive from patients with NSCLC were positive for the T790M mutation, and two of these samples were also positive for the L858R mutation. The simultaneous occurrence of mutations may be significant in a clinical setting, suggesting a possible specific disease pattern or subtype within the studied group. Additionally, all four control samples from healthy donors tested negative, demonstrating the kit’s reliability in correctly identifying true negative samples within the sample set described above. Collectively, these initial findings suggest that the NEAT Liquid Biopsy Kit will enable accurate cancer mutation detection in NSCLC samples for research use.



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