吖啶橙共振光散射法测定痕量脱氧核糖核酸

Determination of Deoxyribonucleic Acids at Nanogram Levels with Acridine Orange by a Resonance Light-Scattering Technique

研究了三环杂芳香类染料吖啶橙 (AO)与DNA作用的共振光散射光谱 ,在 pH11.5～12.5的范围内 ,加入DNA导致吖啶橙共振光散射增强 ,在339nm处 ,存在一共振光散射增强峰 ,其强度与DNA浓度呈线性关系 ,据此建立了一种测定DNA的共振光散射新方法。对于ctDNA ,方法的线性范围为14.3～1000μg/L ,检出限为2.86μg/L,RSD为3.6 %;对于fsDNA ,方法的线性范围为24.0～1250μg/L ,检出限为4.78μg/L,RSD为6.0%。

The resonance light scattering(RLS)spectra of the Acridine Orange-DNA conjugate were studˉied.The presence of DNA greatly enhanced the RLS intensity of Acridine Orange(AO)in a pH range of11.5～12.5.A resonance light scattering peak at339nmwas found,and the enhanced RLS intensity at this wavelength was proportional to the concentration of DNA.Thus,a newRLS method was established for the deˉtermination of DNA.The linear ranges for calf thymus DNA(ctDNA)and fish spermDNA(fsDNA)were found to be14.3～1000μg/Land24.0～1250μg/Lwith detection limits of2.86μg/Land4.78μg/L,respecˉtively.The precision for ctDNA at500μg/L was3.6%and that for fsDNA at the same level was6.0%.Four synthetic samples were determined satisfactorily.