Shigenaga M K, Aboujaoude E N, Chen Q, Ames B N
Department of Molecular and Cell Biology, University of California at Berkeley 94720.
Methods Enzymol. 1994;234:16-33. doi: 10.1016/0076-6879(94)34073-0.
High-performance liquid chromatography with electrochemical detection is a highly sensitive and selective method for detecting oxo8dG and oxo8Gua, biomarkers of oxidative DNA damage. When employed together with the DNA isolation and monoclonal antibody-based immunoaffinity purification methods described, oxo8dG and oxo8Gua in DNA and urine can be readily detected and quantitated, offering a powerful approach for assessing oxidative DNA damage in vivo. Application of the technique to the detection of oxo8dG from DNA permits quantitation of the steady-state levels of this oxidatively modified deoxynucleoside and overcomes the detection problems associated with the extremely low levels present in DNA. In addition, the selectivity gained by this detection method eliminates the problem of separating the signal for oxo8dG from those of normal deoxynucleosides. The quantitation of oxo8dG and oxo8Gua in biological fluids is noninvasive and complements the measurement of oxo8dG in DNA by estimating the rate of oxidative DNA damage occurring within the body or in a population of cells. This analytical approach may allow one to estimate oxidative DNA damage in an animal or individual exposed to prooxidant conditions associated with lifestyle, genetic predisposition, degenerative diseases, or environmental toxins. Furthermore, these assays may allow one to assess the potentially beneficial effects of intervention strategies that protect DNA from such damage.
高效液相色谱电化学检测法是检测8-氧代脱氧鸟苷（oxo8dG）和8-氧代鸟嘌呤（oxo8Gua）的高灵敏度、高选择性方法，这两种物质是氧化性DNA损伤的生物标志物。当与所述的DNA分离及基于单克隆抗体的免疫亲和纯化方法一起使用时，DNA和尿液中的oxo8dG和oxo8Gua能够很容易地被检测和定量，为评估体内氧化性DNA损伤提供了一种有力的方法。将该技术应用于从DNA中检测oxo8dG，可以对这种氧化修饰的脱氧核苷的稳态水平进行定量，并克服了与DNA中极低水平相关的检测问题。此外，这种检测方法所获得的选择性消除了将oxo8dG的信号与正常脱氧核苷的信号分离的问题。生物体液中oxo8dG和oxo8Gua的定量检测是非侵入性的，通过估计体内或细胞群体中发生的氧化性DNA损伤速率，对DNA中oxo8dG的测量起到补充作用。这种分析方法可以让人们估计在暴露于与生活方式、遗传易感性、退行性疾病或环境毒素相关的促氧化条件下的动物或个体的氧化性DNA损伤。此外，这些检测方法可以让人们评估保护DNA免受此类损伤的干预策略的潜在有益效果。
