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基于共振光散射技术开发一种使用氯化四苯基卟啉铁的灵敏且快速的核酸检测方法。

Development of a sensitive and rapid nucleic acid assay with tetraphenyl porphyrinatoiron chloride by a resonance light scattering technique.

作者信息

Chen Zhanguang, Liu Jinbin, Zhu Li, Ding Weifeng, Han Yali

机构信息

Department of Chemistry, Shantou University, Shantou 515063, People's Republic of China.

出版信息

Luminescence. 2007 Sep-Oct;22(5):493-500. doi: 10.1002/bio.991.

Abstract

Based on the interaction between nucleic acids and tetraphenyl porphyrinatoiron chloride (FeTPPCl), a novel method for the determination of nucleic acids at the nanogram level has been developed. Under the optimum conditions, the weak resonance light scattering (RLS) intensity of FeTPPCl was greatly enhanced by nucleic acids and the enhanced RLS intensity was proportional to the concentration of nucleic acids in the range 0.02-2.8 microg/mL for calf thymus DNA, 0.05-3.3 microg/mL for fish sperm DNA and 0.07-4.5 microg/mL for yeast RNA. The detection limits (3sigma) were 2.9 ng/mL for calf thymus DNA, 3.9 ng/mL for fish sperm DNA and 5.2 ng/mL for yeast RNA. Almost no interference could be observed from proteins, nucleosides and most of the metal ions. The proposed method showed good reliability, sensitivity, rapidity and reproducibility when applied to the determination of nucleic acids in synthetic and biochemical samples. The time savings make this method suitable for large routine analyses.

摘要

基于核酸与四苯基氯化卟啉铁(FeTPPCl)之间的相互作用,开发了一种用于测定纳克级核酸的新方法。在最佳条件下,核酸极大地增强了FeTPPCl的弱共振光散射(RLS)强度,增强后的RLS强度与核酸浓度成正比,小牛胸腺DNA的浓度范围为0.02 - 2.8μg/mL,鱼精DNA的浓度范围为0.05 - 3.3μg/mL,酵母RNA的浓度范围为0.07 - 4.5μg/mL。小牛胸腺DNA的检测限(3σ)为2.9 ng/mL,鱼精DNA的检测限为3.9 ng/mL,酵母RNA的检测限为5.2 ng/mL。蛋白质、核苷和大多数金属离子几乎不产生干扰。该方法应用于合成样品和生化样品中核酸的测定时,具有良好的可靠性、灵敏度、快速性和重现性。节省时间使得该方法适用于大量的常规分析。

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