A new quantitative method for circulating DNA level in human serum by capillary zone electrophoresis with laser-induced fluorescence detection.
We present a method for the quantification of circulating DNA in serum by capillary zone electrophoresis (CZE) with laser-induced fluorescence detection (LIF). The serum was digested by proteinase to release free DNA. SYBR Gold was utilized as DNA intercalating dye, fluorescein as internal standard (ISTD). CZE-LIF was applied for the separation and quantification of total circulating DNA. Good linearity (R = 0.9992) in the low range of DNA concentrations (0.5-40 ng/mL) and a detection limit of 0.5 ng/mL for DNA (S/N = 3) were obtained. Our data demonstrated that CZE-LIF system has a good linearity with excellent sensitivity and satisfactory reproducibility in the quantification of circulating DNA in serum. This method was successfully used for the quantification of circulating DNA levels in serum. We observed that the circulating DNA levels in certain cancer patients were significantly higher than that in healthy individuals. Compared to current methods, our protocol does not need the extraction of DNA from serum. Our preliminary results have illustrated that CZE-LIF system is simple, rapid, and sensitive, and it is well suitable for large-scale quantification of circulating DNA levels in clinical diagnosis.
Zhang
Peng
P
Department of Chemistry, Shanghai Jiaotong University, Shanghai, PR China.
Ren
Jicun
J
Shen
Zuojun
Z
D000328:Adult; D004396:Coloring Agents / Q000737:chemistry*; D004247:DNA / Q000032:analysis* / Q000097:blood; D019075:Electrophoresis, Capillary / Q000379:methods*; D005260:Female; D005453:Fluorescence; D006801:Humans; D008297:Male; D044967:Serum / Q000737:chemistry*
D004396:Coloring Agents; D004247:DNA