基于荧光染料和氧化石墨烯同步荧光法同时检测多种流感病毒亚型
发布时间：2018-07-27 20:36
【摘要】：分别采用三种不同发射波长的荧光染料,通过共价偶联的方式分别标记三种流感病毒亚型H1N1、H5N1和H9N2的抗体,再利用氧化石墨烯猝灭所标记染料的荧光。当将荧光标记抗体和氧化石墨烯一并加入到流感病毒溶液中时,由于抗原和抗体之间的特异性相互作用,病毒会和抗体作用而使得氧化石墨烯远离荧光染料,染料的荧光得以恢复。通过恢复的荧光发射波长位置和荧光强度,可以定性和定量检测三种不同的流感病毒亚型。在最佳实验条件下,对三种流感病毒亚型H1N1、H5N1和H9N2进行同时检测,H1N1的检出限为0.48ng/mL,线性范围为1~18ng/mL;H5N1的检出限为0.46ng/mL,线性范围为1~18.5ng/mL;H9N2的检出限为0.42ng/mL,线性范围为1~16ng/mL。该方法具有较好的稳定性、重现性和灵敏度,可实现多个流感病毒亚型的分型和同时检测。
[Abstract]:Three kinds of fluorescent dyes with different emission wavelengths were used to label the antibodies against H5N1 and H9N2 by covalent coupling, and then the fluorescence of the dyes labeled by graphene oxide was quenched by graphene oxide. When the fluorescent labeled antibody and graphene oxide are added to the influenza virus solution, because of the specific interaction between the antigen and the antibody, the virus acts with the antibody to keep graphene oxide away from the fluorescent dye. The fluorescence of the dye was restored. Three different influenza virus subtypes can be detected qualitatively and quantitatively by fluorescence emission wavelength and fluorescence intensity. Under the best experimental conditions, the detection limit of H _ (1) N _ (1) H _ 1 and H9N2 for simultaneous detection of H _ (1) N _ (1) was 0.48 ng 路mL ~ (-1), the linear range for H5N1 was 0.46 ng / mL ~ (-1), and the linear range was 0.42 ng / m ~ (-1) for 1n ~ (18) ng 路m ~ (-1) 路m ~ (-1) N ~ (-1) H _ 9N _ 2, and the linear range was 1 ~ (16) ng 路m ~ (-1) 路L ~ (-1) for H _ (1) N _ (1) H _ (1) N ~ (-1) H _ 9N _ 2, respectively. The method has good stability, reproducibility and sensitivity, and can be used for typing and simultaneous detection of multiple influenza virus subtypes.
【作者单位】： 华中农业大学理学院化学系;生物医学分析化学教育部重点实验室武汉大学化学与分子科学学院;
【基金】：国家自然科学基金(No.21305049;21475101)
【分类号】：R511.7;O657.3
，
本文编号：2149070
[Abstract]:Three kinds of fluorescent dyes with different emission wavelengths were used to label the antibodies against H5N1 and H9N2 by covalent coupling, and then the fluorescence of the dyes labeled by graphene oxide was quenched by graphene oxide. When the fluorescent labeled antibody and graphene oxide are added to the influenza virus solution, because of the specific interaction between the antigen and the antibody, the virus acts with the antibody to keep graphene oxide away from the fluorescent dye. The fluorescence of the dye was restored. Three different influenza virus subtypes can be detected qualitatively and quantitatively by fluorescence emission wavelength and fluorescence intensity. Under the best experimental conditions, the detection limit of H _ (1) N _ (1) H _ 1 and H9N2 for simultaneous detection of H _ (1) N _ (1) was 0.48 ng 路mL ~ (-1), the linear range for H5N1 was 0.46 ng / mL ~ (-1), and the linear range was 0.42 ng / m ~ (-1) for 1n ~ (18) ng 路m ~ (-1) 路m ~ (-1) N ~ (-1) H _ 9N _ 2, and the linear range was 1 ~ (16) ng 路m ~ (-1) 路L ~ (-1) for H _ (1) N _ (1) H _ (1) N ~ (-1) H _ 9N _ 2, respectively. The method has good stability, reproducibility and sensitivity, and can be used for typing and simultaneous detection of multiple influenza virus subtypes.
【作者单位】： 华中农业大学理学院化学系;生物医学分析化学教育部重点实验室武汉大学化学与分子科学学院;
【基金】：国家自然科学基金(No.21305049;21475101)
【分类号】：R511.7;O657.3
，
本文编号：2149070
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