Development of an up-converting phosphor technology-based 10-channel lateral flow assay for profiling antibodies against Yersinia pestis
Development of an up-converting phosphor technology-based 10-channel lateral flow assay for profiling antibodies against Yersinia pestis
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DOI：
10.1016/j.mimet.2010.08.005
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发表时间：
2010-11-01
影响因子：
2.2
通讯作者：
Zhou, Lei
中科院分区：
文献类型：
--
作者：
Hong, Wenyan;Huang, Lihua;Zhou, Lei
在这项研究中，开发了一种基于10通道上转换荧光技术的侧向流（TC-UPT-LF）测定法来分析鼠疫耶尔森菌抗体。10例表达Y.将鼠疫蛋白与上转换荧光颗粒共价结合，以开发双抗原夹心免疫层析条，以检测相应的抗体。在逐一优化后，将每个条带整合到TC-UPT-LF盘中，用于同时检测不同的抗体。还开发了扫描生物传感器以获得结果。使用标准样品和鼠疫猴血清样品评价了TC-UPT-LF测定的性能。用TC-UPT-LF法检测了51份患者血清。TC-UPT-LF压片在37 ℃下可稳定10天，平均CV为10.3%。其灵敏度和定性结果与ELISA法相当。其对不同抗体检测的线性拟合决定系数（R-2）在0.93 ~ 0.99之间。除F1抗体外，LcrV和YopD抗体在猴血清中的阳性率分别为100%（13/13）和92%（12/13），在病人血清中的阳性率分别为86.3%（44/51）和66.7%（34/51）。结果表明，TC-UPT-LF检测方法已成功建立，LcrV和YopD可作为鼠疫的潜在诊断标志物。(C)2010 Elsevier B. V.保留所有权利。
In this study, a 10-channel up-converting phosphor technology-based lateral flow (TC-UPT-LF) assay was developed to profile antibodies against Yersinia pestis. Ten expressed Y. pestis proteins were covalently conjugated with an up-converting phosphor particle to develop double-antigen sandwich immunochromatographic strips to detect corresponding antibodies. After optimization one by one, each strip was integrated into a TC-UPT-LF disc for simultaneously detection of different antibodies. A scanning biosensor was also developed to acquire the results. The performance of the TC-UPT-LF assay was evaluated by using standard samples and plague monkey serum samples. Fifty-one patient serum samples were detected by the TC-UPT-LF assay. The TC-UPT-LF disc could be stable for 10 days at 37 degrees C with an average CV of 10.3%. Its sensitivity and qualitative results are comparable to those of ELISA. Its linearity fitting coefficient of determination (R-2) for different antibody detection is between 0.93 and 0.99. Besides F1 antibody, the LcrV and YopD antibodies also showed higher positive ratio than the other seven antibodies, as 100% (13/13) and 92% (12/13) in monkey sera and 86.3% (44/51) and 66.7% (34/51) in patient sera, respectively. It is suggested that the TC-UPT-LF assay has been successfully developed for multi-detection and LcrV and YopD can be the potential diagnostic markers of the plague. (C) 2010 Elsevier B.V. All rights reserved.