【摘要】：分別采用三種不同發(fā)射波長的熒光染料,通過共價(jià)偶聯(lián)的方式分別標(biāo)記三種流感病毒亞型H1N1、H5N1和H9N2的抗體,再利用氧化石墨烯猝滅所標(biāo)記染料的熒光。當(dāng)將熒光標(biāo)記抗體和氧化石墨烯一并加入到流感病毒溶液中時(shí),由于抗原和抗體之間的特異性相互作用,病毒會(huì)和抗體作用而使得氧化石墨烯遠(yuǎn)離熒光染料,染料的熒光得以恢復(fù)。通過恢復(fù)的熒光發(fā)射波長位置和熒光強(qiáng)度,可以定性和定量檢測(cè)三種不同的流感病毒亞型。在最佳實(shí)驗(yàn)條件下,對(duì)三種流感病毒亞型H1N1、H5N1和H9N2進(jìn)行同時(shí)檢測(cè),H1N1的檢出限為0.48ng/mL,線性范圍為1~18ng/mL;H5N1的檢出限為0.46ng/mL,線性范圍為1~18.5ng/mL;H9N2的檢出限為0.42ng/mL,線性范圍為1~16ng/mL。該方法具有較好的穩(wěn)定性、重現(xiàn)性和靈敏度,可實(shí)現(xiàn)多個(gè)流感病毒亞型的分型和同時(shí)檢測(cè)。
[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.
【作者單位】： 華中農(nóng)業(yè)大學(xué)理學(xué)院化學(xué)系;生物醫(yī)學(xué)分析化學(xué)教育部重點(diǎn)實(shí)驗(yàn)室武漢大學(xué)化學(xué)與分子科學(xué)學(xué)院;
【基金】：國家自然科學(xué)基金(No.21305049;21475101)
【分類號(hào)】：R511.7;O657.3
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本文編號(hào)：2149070