Quantum Dots Labeled Antibody as a Fluorescence Probe for Hantavirus Fluoroimmunoassay
Quantum Dots Labeled Antibody as a Fluorescence Probe for Hantavirus Fluoroimmunoassay
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量子点标记抗体作为汉坦病毒荧光免疫分析的荧光探针
DOI：
10.1166/jnn.2016.11300
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发表时间：
2016-07
影响因子：
--
通讯作者：
姚苹苹等
中科院分区：
文献类型：
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作者：
姚苹苹等
量子点现在被广泛用于免疫检测目的。我们开发了一种实用、稳定的量子点标记羊抗人IgG试剂，作为高荧光强度的光稳定标记物，用于有效的抗体检测。有效标记所需的最佳反应时间、pH值和羊抗人IgG浓度分别为2 h、6.0和20 μg/mL。结合间接免疫荧光技术，我们建立了一种以量子点代替化学荧光染料的汉坦病毒（HV）免疫检测系统。该免疫检测方案快速、特异、灵敏，检测范围为5 ng/mL ~ 10 μg/mL，与20份B型肝炎病毒（HBV）、单纯疱疹病毒1型（HSV 1）和丙型肝炎病毒（HCV）阳性标本无交叉反应。对HV和HBV免疫检测系统的优化响应时间显示，抗体-抗原结合反应在30 min内达到平衡，而抗体和量子点标记的羊抗人IgG结合反应在10 min内达到平衡，整个免疫检测过程约40 min完成。以PCR为金标准，对207份临床血清样品进行了QDs和酶联免疫吸附试验（ELISA）检测方法比较。QDs免疫检测的敏感性为96%，特异性为98.7%，约登指数为94.7%。比较量子点和ELISA免疫检测系统的匹配卡方检验结果显示，两种方法之间没有显着差异。量子点-抗体-抗原复合物具有较长的荧光寿命，使其成为监测或审查结果的相对简单的过程。这种新方法有可能用于人类HV感染的常规检测。
Quantum dots are now widely used for immune detection purposes. We developed a practical and robust quantum dots labeled goat anti-human IgG reagent as a high fluorescent intensity photostable marker for effective antibody detection. The optimum reaction time, pH, and concentration of goat anti-human IgG required for effective labeling was 2 h, 6.0, and 20 μg/mL, respectively. Combined with an indirect immunofluorescence assay, we developed an immune detection system for Hantavirus (HV) using quantum dots instead of chemical fluorochromes. This immune detection scheme was rapid, specific, and sensitive with a detection range of 5 ng/mL–10 μg/mL, and no cross-reaction with 20 samples positive for hepatitis B virus (HBV), herpes simplex virus 1, and hepatitis C virus. The optimized response time of the immune detection system for HV and HBV revealed that the antibody-antigen binding reaction equilibrated within 30 min, while the antibody and QDs labeled goat anti-human IgG combination equilibrated in 10 min. The whole immune detection process took around 40 min to complete. PCR was used as the gold standard for comparing the QDs and enzyme linked immunosorbent assay (ELISA) detection methods using 207 clinical serum samples. The sensitivity of QDs immune detection was 96%, its specificity was 98.7%, while the Youden index was 94.7%. The results of the matched chi-square test used to compare the QDs and ELISA immune detection systems revealed no significant difference between the two methods. The QDs-antibody-antigen complex had a long fluorescence lifetime, making it a relatively straightforward process to monitor or review the results. This new method has potential for routine detection of HV infection in humans.