用四氨基铝酞菁共振光散射技术测定纳克级核酸

Determination of Nucleic Acids at Nanogram Levels with Tetra-substituted Amino Aluminum Phthalocyanine by a Resonance Light-scattering Technique

痕量核酸对四氨基铝酞菁的共振散射光产生增强作用 ,且增强程度与核酸浓度之间有良好的线性关系 .据此建立了测定核酸的高灵敏共振散射光增强分析方法 .在 p H=6.0、最大散射波长 4 0 0 nm处 ,测定小牛胸腺 DNA( CT DNA)、鲑鱼精子 DNA( SM DNA)和酵母 RNA( Yeast RNA)的线性范围分别是0～ 2 50 ng/ m L,0～ 2 0 0 ng/ m L和 0～ 4 0 0 ng/ m L,检测限分别为 1 .4 ng/ m L,1 .4 ng/ m L和 2 .7ng/ m L.该法简单 ,灵敏度高 ,用于实际样品中核酸含量的测定 。

This is the first report of the determination of nucleic acids at nanogram levels based on their enhanced effects on the resonance light scattering of TAAlPc at 400 nm. Under optimal conditions, the linear ranges of the calibration curves were 0—250 0 ng/mL for Calf thymus DNA(CT DNA), 0—200.0 ng/mL for Salmon sperm DNA(SM DNA) and 0—400 0 ng/mL for Yeast RNA. The detection limits were 1.4 ng/mL for both CT DNA and SM DNA, 2.7 ng/mL for Yeast RNA. The relative standard deviation for the determination of 100.0 ng/mL CT DNA was 2.1%. The measurement can be made easily on a common spectrofluorimeter. The reaction between TAAlPc and nucleic acids is completed in 2 min and the scattered light signal is stable for at least 1 h. The interference from some interesting co existing substances with the determination of DNA was also examined. The method was applied to the determination of DNA in golden staphylococcus DNA sample, and the results were satisfactory.