Digital PCR detection of EGFR somatic mutations in non-small-cell lung cancer formalin fixed paraffin embedded samples - PubMed
Source : https://pubmed.ncbi.nlm.nih.gov/34089806/
doi: 10.1016/j.mcp.2021.101745. Online ahead of print. 1 Laboratório de Genética Molecular Humana, Universidade Federal Do Paraná, Curitiba, Brazil. 2 Laboratório de Genética Molecular Humana, Universidade Federal Do Paraná, Curitiba, Brazil; Laboratório Genoprimer, Diagnóstico Molecular e Medicina Personalizada, Curitiba, Brazil. 3 Laboratório Genoprimer, Diagnóstico Molecular e Medicina Personalizada, Curitiba, Brazil.
Key Points
• Digital PCR (dPCR) could replace real time PCR and Sanger sequencing in the detection/quantification of rare mutations, such as EGFR. In the current study, researchers compared the performance of dPCR and castPCR in 57 NSCL formalin fixed paraffin embedded samples and 10 lung cancer-free formalin fixed paraffin embedded samples. They found that dPCR vs. castPCR had a higher sensitivity and specificity when screening EGFR mutations in NSCLC biopsies.
• “In this work, we found that dPCR was able to detect the most frequent EGFR mutations DEL19 and in more than 50% of the samples analyzed. In contrast, only 4 samples were detected with DEL19 by castPCR, evidencing the greater sensitivity of dPCR in detecting this deletion. For , two samples were detected by castPCR, indicating a trend for the best performance of dPCR. The use of highly sensitive assays for the detection of the p.T790 M mutation has been reported to yield false positive results, especially in samples with reduced DNA content, making it difficult to distinguish false from true positives. However, we found no differences in the performance of dPCR and castPCR for p.T790 M detection,” the authors wrote.
• The authors noted that although tissue biopsy is the current gold standard, it can require re-biopsy. Furthermore, tissue specimens can only be subjected to a limited number of tests.