Comparison of three commonly used PCR‐based techniques to analyze MSI status in sporadic colorectal cancer

Abstract

Several retrospective studies have shown that a high level of microsatellite instability (MSI-H) is an important prognostic factor of a more favorable outcome in stage II and III colorectal cancer (CRC) patients. In this study, three commonly used polymerase chain reaction (PCR)-based MSI analysis techniques were compared (polyacrylamide gel electrophoresis followed by silver-staining [SSPAGE], fluorescence capillary electrophoresis [FCE], and denaturing high-performance liquid chromatography [DHPLC]) on a limited group of CRC patients, to identify the most optimal detection technique. Pathology blocks of 26 CRC patients were subjected to microdissection and the Bethesda reference panel was used for MSI analysis. Considering the samples analyzed by both SSPAGE and FCE, 8.7% were MSI-H, 8.7% were MSI-L, and 82.6% were MSS using SSPAGE. FCE resulted in 16% MSI-H, 4% MSI-L, and 80% MSS. Due to difficulties in analyzing the dinucleotide markers on DHPLC, we only analyzed the mononucleotide markers with this technique. The results were 100% concordant to those obtained by FCE. SSPAGE is time consuming, subjective, and less user-friendly and interpretable. DHPLC was not feasible due to interpretation difficulties for the dinucleotide markers. We recommend the use of FCE to analyze MSI status. This technique is sensitive, reproducible, user-friendly and leads to easy interpretation and high-throughput.