EGFR PCR Test for NSCLC Diagnosis

The EGFR PCR test (cobas EGFR Mutation Test, Roche Molecular Systems, Inc, Branchburg, NJ, USA) is a CE-IVD marked multiplex allele-specific PCR-based assay designed to detect 41 mutations in exons 18, 19, 20, and 21 in FFPET specimens of human NSCLC.[28] (link) DNA is isolated using the cobas DNA Sample Preparation Kit (Roche Molecular Systems, Branchburg, NJ). [29] A minimum of 150 ng of genomic DNA is required for PCR amplification, which can typically be isolated from a single 5 µm FFPET section. The EGFR PCR test software version used in this study was designed to detect 29 deletions in exon 19 and 2 L858R variants in exon 21. Macrodissection is only recommended if tumor content is less than 10%; laser capture microdissection is not required. The EGFR PCR test was performed per manufacturer's package insert and results were automatically analyzed and reported. The limit of detection has been validated to 5% mutant alleles. The workflow from DNA isolation to results reporting can be performed in one 8 hour period.[27]