Preliminary Study on the Identification of BRAF(V600E) Mutation in Colorectal Cancer by Near-Infrared Spectroscopy

INTRODUCTION: In metastatic colorectal cancer (mCRC), the B-type Raf kinase (BRAF)(V600E) mutation is a molecular biomarker of poor prognosis and is of great importance to drug target. Currently, the commonly used methods for detecting BRAF(V600E) mutation include immunohistochemistry (IHC) and gene sequencing, but both present certain limitations. Near-infrared (NIR) spectroscopy is a spectroscopy technology that takes advantage of the electromagnetic wavelength between visible light and mid-infrared light. METHODS: IHC was used to detect the expression of BRAF(V600E) protein with the BRAF(V600E) (VE1) antibody in 42 cases of paraffin-embedded (FFPE) mCRC tissue sections. The NIR-discriminant analysis model (NIRS-DA) was established using 6 cases of wild-type and 6 cases of mutant-type BRAF specimens. RESULTS: IHC detection results revealed 13 cases of weakly positive (+), 1 case of moderately positive (++), and 28 cases of negative (−) CRC. Compared with the next-generation sequencing (NGS) results, the positive rate was 66.7%. The classification accuracy of calibration (CAC) was 100% compared with the results of NGS, demonstrating that the BRAF(V600E) mutant NIRS-DA model, verified by 2 cases of wild-type and 2 cases of mutant-type CRC samples was established. The NIRS-DA model was used to predict gene mutation in the CRC samples, 7 cases were positive (+), and 35 cases were negative (−), and the classification accuracy of prediction (CAP) was 83.3% (35/42). DISCUSSION: The NIRS-DA model-predicted results were in high agreement with the detection results of NGS, and the difference in IHC is not statistically significant (P>0.05). However, this study is a preliminary discussion on a methodology due to its small sample size.