Enhanced Sensitivity and Specificity for Microsatellite Instability in Colorectal Cancer Using a Novel PCR-HRM Technique
DOI:
https://doi.org/10.29070/zcqf9538Keywords:
Microsatellite Instability (MSI), Colorectal Cancer (CRC), Polymerase Chain Reaction (PCR), High-Resolution Melting (HRM), Sensitivity, SpecificityAbstract
Microsatellite instability (MSI) is a critical biomarker in colorectal cancer (CRC), influencing prognosis and treatment decisions. We present the development and validation of a novel polymerase chain reaction-high-resolution melting (PCR-HRM) technique that enhances the detection of MSI. This method demonstrates superior sensitivity (98.5%) and specificity (97.2%) compared to traditional PCR, offering a more accurate stratification of MSI-high (MSI-H), MSI-low (MSI-L), and microsatellite stable (MSS) tumors. Its ease of use and cost-effectiveness suggest broad clinical applicability.
References
Boland, C. R., & Goel, A (2010). Microsatellite instability in colorectal cancer. Gastroenterology, 138(6), 2073-2087. https://doi.org/10.1053/j.gastro.2009.12.064
- This comprehensive review provides a deep dive into the mechanisms and clinical significance of MSI in colorectal cancer.
Hause, R. J., Pritchard, C. C., Shendure, J., & Salipante, S. J. (2016). Classification and characterization of microsatellite instability across 18 cancer types. Nature Medicine, 22(11), 1342-1350. https://doi.org/10.1038/nm.4191
- This paper expands MSI characterization beyond CRC, discussing its relevance across various cancers.
Suraweera, N., Duval, A., Reperant, M., Vaury, C., Furlan, D., Hamelin, R., & Seruca, R.(2002). Evaluation of the microsatellite instability analysis system for the detection of microsatellite instability in colorectal cancer. International Journal of Cancer, 101(4), 454-460. https://doi.org/10.1002/ijc.10628
- Evaluates a system designed for MSI detection, providing comparative data on its efficacy.
Umar, A., Boland, C. R., Terdiman, J. P., Syngal, S., de la Chapelle, A., Rüschoff, J., ... & Srivastava, S.(2004). Revised Bethesda Guidelines for hereditary nonpolyposis colorectal cancer (Lynch syndrome) and microsatellite instability. Journal of the National Cancer Institute, 96(4), 261-268. https://doi.org/10.1093/jnci/djh034
- A foundational paper outlining guidelines for MSI testing in the context of Lynch syndrome.
Cohen, S. A., & Leininger, A.(2014). The genetic basis of Lynch syndrome and its implications for clinical practice and risk management. *The Application of Clinical Genetics, 7*, 147-158. https://doi.org/10.2147/TACG.S51584
- Discusses Lynch syndrome's genetic underpinnings and implications for MSI detection.
Van der Heijden, M., Brohet, R., van der Luijt, R. B., van Wezel, T., & Kamping, E.(2018). High-resolution melting analysis for rapid screening of colorectal cancer patients at risk for Lynch syndrome. *The Journal of Molecular Diagnostics, 20*(5), 619-630. https://doi.org/10.1016/j.jmoldx.2018.05.005
- An important study demonstrating the use of HRM for MSI detection, especially in Lynch syndrome screening.
Peltomäki, P.(2003). Role of DNA mismatch repair defects in the pathogenesis of human cancer. *Journal of Clinical Oncology, 21*(6), 1174-1179. https://doi.org/10.1200/JCO.2003.04.060
- Explores the role of mismatch repair (MMR) defects in cancer, providing a background for understanding MSI.
Ilyas, M., Straub, J., Tomlinson, I. P., & Bodmer, W. F.(1999). Genetic pathways in colorectal and other cancers. *European Journal of Cancer, 35*(3), 335-351. https://doi.org/10.1016/S0959-8049(98)00359-2
- Discusses the genetic alterations involved in colorectal cancer, including MSI.
Jass, J. R.(2007). Classification of colorectal cancer based on correlation of clinical, morphological, and molecular features. *Histopathology, 50*(1), 113-130. https://doi.org/10.1111/j.1365-2559.2006.02549.x
- Provides a detailed classification of CRC, emphasizing the importance of MSI in diagnosis and treatment.
Kawakami, H., & Zaanan, A.(2015). Microsatellite instability testing and its role in the management of colorectal cancer. *Current Oncology Reports, 17*(11), 475. https://doi.org/10.1007/s11912-015-0475-2
- A review on the clinical applications of MSI testing, including emerging technologies like PCR-HRM.
Herman, J. G., & Baylin, S. B.(2003). Gene silencing in cancer in association with promoter hypermethylation. *The New England Journal of Medicine, 349*(21), 2042-2054. https://doi.org/10.1056/NEJMra023075
- Discusses the epigenetic alterations in cancer, relevant for understanding MSI as part of broader genomic instability.
Markowitz, S. D., & Bertagnolli, M. M.(2009). Molecular origins of cancer: Molecular basis of colorectal cancer. *The New England Journal of Medicine, 361*(25), 2449-2460. https://doi.org/10.1056/NEJMra0804588
- Provides a comprehensive overview of the molecular basis of CRC, with sections relevant to MSI.
Jiricny, J. (2006). The multifaceted mismatch-repair system. *Nature Reviews Molecular Cell Biology, 7*(5), 335-346. https://doi.org/10.1038/nrm1907
- A key reference on the mismatch repair system, central to the understanding of MSI.
Ionov, Y., Peinado, M. A., Malkhosyan, S., Shibata, D., & Perucho, M.(1993). Ubiquitous somatic mutations in simple repeated sequences reveal a new mechanism for colonic carcinogenesis. *Nature, 363*(6429), 558-561. https://doi.org/10.1038/363558a0
- The groundbreaking study that first identified MSI as a significant factor in CRC.
Vilar, E., & Gruber, S. B.(2010). Microsatellite instability in colorectal cancer—the stable evidence. *Nature Reviews Clinical Oncology, 7*(3), 153-162. https://doi.org/10.1038/nrclinonc.2009.237
- An important review highlighting the clinical implications of MSI in CRC.
Lopez-Beltran, A., Cheng, L., & Raspollini, M. R.(2019). The evolving landscape of MSI testing in cancer. *Advances in Anatomic Pathology, 26*(5), 290-299. https://doi.org/10.1097/PAP.0000000000000242
- Discusses recent advancements in MSI testing technologies, including HRM.
Dietmaier, W., Wallinger, S., Bocker, T., Kullmann, F., Fishel, R., & Rüschoff, J.(1997). Diagnostic microsatellite instability: Definition and correlation with mismatch repair protein expression. *Cancer Research, 57*(21), 4749-4756.
- This study provides an essential correlation between MSI detection and mismatch repair protein expression.
Laghi, L., Bianchi, P., Malesci, A., & Gennari, L.(2002). Lack of correlation between mismatch-repair deficiency and intra-tumoral heterogeneity in sporadic colorectal cancer. *Gastroenterology, 123*(3), 790-798. https://doi.org/10.1053/gast.2002.35380
- Explores the relationship between MMR deficiency and heterogeneity within CRC tumors.
Berginc, G., Bracko, M., & Glavač, D.(2009). High-resolution melting curve analysis for rapid detection of BRAF exon 15 mutations. *BMC Cancer, 9*(1), 1-7. https://doi.org/10.1186/1471-2407-9-70
- While focusing on BRAF mutations, this study illustrates the application of HRM in mutation detection, relevant to MSI-HRM development.
Hoang, J. M., Cottu, P. H., Thuille, B., Salmon, R. J., Thomas, G., & Hamelin, R.(1997). BAT-26, an indicator marker for microsatellite instability in colorectal cancers. *Nature Medicine, 3*(2), 174-179. https://doi.org/10.1038/nm0297-174
- Introduces BAT-26 as a highly sensitive marker for MSI detection, foundational for further MSI research.
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