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首页> 外文期刊>Analytical chemistry >Spectrally and Spatially Multiplexed Serological Array-in-Well Assay Utilizing Two-Color Upconversion Luminescence Imaging
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Spectrally and Spatially Multiplexed Serological Array-in-Well Assay Utilizing Two-Color Upconversion Luminescence Imaging

机译：光谱和空间多路复用血清学阵列在井中利用两色上转换发光成像

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摘要

We demonstrate a simple dual-mode multiplexed array-in-well immunoassay for simultaneous classification and detection of serum IgG and IgM antibodies against influenza A and human adenoviruses based on the color and position of the upconversion luminescence on the array. Biotinylated influenza A/H1N1 and A/H5N1 as well as adenovirus serotype 2 and 5 hexon antigens along with positive and negative controls were printed in an array format onto the bottom of streptavidin-coated microtiter wells. The anti-influenza A and antiadenovirus antibodies present in the sample were captured to the array and detected with antihuman antibody-coated upconverting nanophosphors (UCNPs). The green emitting UCNPs (NaYF4:Yb3+,Er3+) coated with antihuman IgG and blue emitting UCNPs (NaYF4:Yb3+,Tm3+) coated with antihuman IgM were used to detect human IgG and IgM antibodies, respectively. The emission of the bound UCNPs was imaged free of autofluorescence with anti-Stokes photoluminescence microwell imager. No spectral cross-talk was observed between green and blue emitting UCNPs. Also the cross-reactivities between UCNP-conjugates and immobilized human IgG and IgM antibodies were negligible. Position of the signal on the array defined the antigen specificity and the antibody class was defined by the color of the upconversion luminescence. This technology could be used for differentiation between acute infection from past infection and immunity. Additionally, the class of the antibody response can be used for the differentiation between primary and secondary infections, hence, facilitating epidemiological seroprevalence studies.
机译：我们展示了一种简单的双模式多孔阵列免疫测定法，可根据上转换发光的颜色和位置在阵列上同时分类和检测针对甲型流感和人类腺病毒的血清IgG和IgM抗体。将生物素化的甲型流感A / H1N1和A / H5N1以及腺病毒血清型2和5六邻体抗原以及阳性和阴性对照以阵列形式印在涂有链霉亲和素的微量滴定孔的底部。样品中存在的抗甲型流感病毒和抗腺病毒抗体被捕获到阵列中，并用抗人抗体涂层的上转换纳米荧光粉（UCNP）检测。涂有抗人IgG的绿色UCNP（NaYF4：Yb3 +，Er3 +）和涂有抗人IgM的蓝色UCNP（NaYF4：Yb3 +，Tm3 +）分别用于检测人IgG和IgM抗体。用抗斯托克斯光致发光微孔成像仪对结合的UCNP的发射进行了无自发荧光成像。在绿色和蓝色发射的UCNP之间未观察到光谱串扰。 UCNP偶联物与固定的人IgG和IgM抗体之间的交叉反应性也可以忽略不计。信号在阵列上的位置定义了抗原特异性，抗体类别由上转换发光的颜色定义。该技术可用于区分急性感染与过去感染和免疫。另外，抗体应答的类别可用于区分原发性感染和继发性感染，因此有助于流行病学血清阳性率研究。

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