| Title: |
A simple, accurate and cost-effective capillary electrophoresis test with computational methods to aid in universal microsatellite instability testing |
| Authors: |
Toh, James; Singh, Puneet; Tangirala, Venkata; Limmer, Alexandra; Spring, Kevin J. (R17176) |
| Publisher Information: |
Switzerland, MDPI |
| Publication Year: |
2021 |
| Collection: |
University of Western Sydney (UWS): Research Direct |
| Subject Terms: |
XXXXXX - Unknown |
| Description: |
Background: Microsatellite instability (MSI) testing is important for the classification of Lynch syndrome, as a prognostic marker and as a guide for adjuvant chemotherapy in colorectal cancer (CRC). The gold standard for determining MSI status has traditionally been fluorescent multiplex polymerase chain reaction (PCR) and capillary gel electrophoresis (CGE). However, its use in the clinical setting has diminished and has been replaced by immunohistochemical (IHC) detection of loss of mismatch repair protein expression due to practicability and cost. The aim of this study was to develop a simple, cost-effective and accurate MSI assay based on CGE. Method: After amplification of microsatellites by polymerase chain reaction (PCR) using the National Cancer Institute (NCI) panel (BAT 25, BAT26, D5S346, D2S123, D17S250) of MSI markers, parallel CGE was utilized to classify colorectal cancers as MSI-H, MSI-L and MSS using the 5200 Fragment Analyzer System. Cell lines and patient cancer specimens were tested. DNA from 56 formalin-fixed paraffin-embedded cancer specimens and matched normal tissue were extracted and CGE was performed. An automated computational algorithm for MSI status determination was also developed. Results: Using the fragment analyser, MSI status was found to be 100% concordant with the known MSI status of cell lines and was 86% and 87% concordant with immunohistochemistry (IHC) from patient cancer specimens using traditional assessment and our MSI scoring system, respectively, for MSI determination. The misclassification rate was mainly attributed to IHC, with only one (1.8%) sampling error attributed to CGE testing. CGE was also able to distinguish MSI-L from MSI-H and MSS, which is not possible with IHC. An MSI score based on total allelic variability that can accurately determine MSI status was also successfully developed. A significant reduction in cost compared with traditional fluorescent multiplex PCR and CGE was achieved with this technique. Conclusions: A simple, cost-effective and ... |
| Document Type: |
article in journal/newspaper |
| File Description: |
print |
| Language: |
English |
| Relation: |
Cells--2073-4409-- Vol. 10 Issue. 6 No. 1401 pp: - |
| DOI: |
10.3390/cells10061401 |
| Availability: |
https://doi.org/10.3390/cells10061401; https://hdl.handle.net/1959.7/uws:67693 |
| Rights: |
© 2021 by the authors. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
| Accession Number: |
edsbas.34A90133 |
| Database: |
BASE |