Application of COLD-PCR for improved detection of EGFR mutations in clinical samples.

• Main Author: Zhang, Zhi Xuan.
• Other Authors: School of Biological Sciences
• Format: Final Year Project
• Language: English
• Published: 2010
• Subjects: DRNTU::Science::Biological sciences::Molecular biology
• Online Access: http://hdl.handle.net/10356/41758
• Institution: Nanyang Technological University

Due to the significance of mutation status of the epidermal growth factor receptor (EGFR) gene in non-small-cell lung cancer (NSCLC) affecting patient response to tyrosine kinase inhibitors (TKI), the more sensitive and robust the method used to detect the mutations, the better. It has been suggested that co-amplification at lower denaturation temperature (COLD)-PCR enhances the amplification of mutant alleles over wild type and hence, may be applied locally to NSCLC patients as a routine test to aid in deciding on the use of TKIs for treatment of NSCLC patients. As such, this project was carried out to find out whether COLD-PCR would pick up low level mutant alleles that were missed by regular PCR. After COLD-PCR and sequencing analysis of 30 samples, this study did not detect any new mutations in the samples when COLD-PCR was used and no significant overall increase in the ratio of mutant to wild type alleles was observed. Thus, based on current data, there is no comparable advantage when using COLD-PCR over regular PCR for mutation detection in routine clinical applications.