Quantitative PCR for analysis of DNA damage induced by PGHS-2 (COX-2) or other factors

摘要

A method to assess DNA damage using quantitative polymerase chain reaction (QPCR) with an internal control with or without modified QPCR primers is disclosed. The method further included quantitation of QPCR products using a chemiluminescent, colorimetric or fluorescent assay and a real-time PCR. The chemiluminescent, colorimetric or fluorescent assay is carried out with modified PCR primers and the real-time PCR is carried out with PCR primers with or without modification. A method to select a suitable PCR replication cycle number is disclosed. This is to avoid polymerization of PCR products after addition of the internal control. A method to analyze DNA damage and mutagenicity induced by chemicals in DNA damage repair-deficient and proficient fibroblast cells with endogenous expression of oxidative stress-inducing proteins, e.g., prostaglandin H₂ synthase form 2 (PGHS-2, also called as COX-2), was disclosed.