Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20, Dongdajie, Fengtai District, Beijing 100071, People's Republic of China.
J Microbiol Methods. 2010 Nov;83(2):133-40. doi: 10.1016/j.mimet.2010.08.005. Epub 2010 Aug 27.
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°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 (R2) 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.
在这项研究中，开发了一种基于 10 通道上转换荧光粉技术的侧向流动（TC-UPT-LF）测定法，用于分析针对鼠疫耶尔森氏菌的抗体。将 10 种表达的鼠疫耶尔森氏菌蛋白与上转换荧光粉颗粒共价偶联，开发双抗原夹心免疫层析条，以检测相应的抗体。经过逐一优化，每条条带都被整合到 TC-UPT-LF 圆盘上，用于同时检测不同的抗体。还开发了一种扫描生物传感器来获取结果。通过使用标准样品和鼠疫猴血清样品评估 TC-UPT-LF 测定法的性能。使用 TC-UPT-LF 测定法检测了 51 份患者血清样本。TC-UPT-LF 盘在 37°C 下可稳定 10 天，平均 CV 为 10.3%。其灵敏度和定性结果与 ELISA 相当。不同抗体检测的线性拟合决定系数（R2）在 0.93 到 0.99 之间。除 F1 抗体外，LcrV 和 YopD 抗体在猴血清中的阳性率也高于其他七种抗体，分别为 100%（13/13）和 92%（12/13），在患者血清中的阳性率分别为 86.3%（44/51）和 66.7%（34/51）。这表明 TC-UPT-LF 测定法已成功用于多检测，LcrV 和 YopD 可能是鼠疫的潜在诊断标志物。
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