Analytical Performance of a Highly Sensitive System to Detect Gene Variants Using Next-Generation Sequencing for Lung Cancer Companion Diagnostics.

The recent increase in the number of molecular targeted agents for lung cancer has led to the demand for the simultaneous testing of multiple genes. Although gene panels using next-generation sequencing (NGS) are ideal, conventional panels require a high tumor content, and biopsy samples often do not meet this requirement. We developed a new NGS panel, called compact panel, characterized by high sensitivity, with detection limits for mutations of 0.14%, 0.20%, 0.48%, 0.24%, and 0.20% for exon 19 deletion, L858R, T790M, V600E, and G12C, respectively. Mutation detection also had a high quantitative ability, with correlation coefficients ranging from 0.966 to 0.992. The threshold for fusion detection was 1%. The panel exhibited good concordance with the approved tests. The identity rates were as follows: positive, 100% (95% confidence interval, 95.5-100); negative, 90.9 (82.2-96.3); positive, 100 (59.0-100); negative, 100 (94.9-100); G12C positive, 100 (92.7-100); G12C negative, 100 (93.0-100); positive, 96.7 (83.8-99.9); negative, 98.4 (97.2-99.2); positive, 100 (66.4-100); negative, 99.0 (94.6-100); positive, 98.0 (89.0-99.9); negative 100 (92.8-100); positive, 93.8 (69.8-100); negative, 100 (94.9-100). The analytical performance showed that the panel could handle various types of biopsy samples obtained by routine clinical practice without requiring strict pathological monitoring, as in the case of conventional NGS panels.