Beijing University of Technology, College of Life Science and Bioengineering, Beijing 100124, China.
Beijing Runbio Biotechnology Development Co., Ltd, Beijing 100106, China.
Biomed Environ Sci. 2020 Mar 20;33(3):174-182. doi: 10.3967/bes2020.024.
To develop a rapid, highly sensitive quantitative method for detecting P24 antigen based on near-infrared fluorescent microsphere immunochromatography.
First, we prepared a lateral flow assay test strip, and labeled the detection antibody using a fluorescent microsphere. Second, we optimized the antibody labeling conditions. Third, we optimized the detection conditions. Fourth, we created a working curve. Fifth, we conducted a methodological assessment of the established fluorescent microsphere immunochromatography method. Sixty-six clinical samples were tested, and we compared the established fluorescent microsphere immunochromatography with the quantitative ELISA method.
According to the working curve, the detection limit of the method is 3.4 pg/mL, and the detection range is 3.4 pg/mL to 10 ng/mL. The average intra-assay recovery was 99.6%, and the Coefficient of Variation (CV) was 5.4%-8.6%; the average inter-assay recovery was 97.3%, and the CV was 8.5%-11%. The detection rate of fluorescent microsphere immunochromatography was higher than ELISA method, and had a good correlation with ELISA.
The P24 antigen quantitative detection method based on near-infrared fluorescent microsphere immunochromatography has the advantages of rapid detection, high sensitivity, and wide detection range; thus, it is suitable for early clinical diagnosis and continuous monitoring of AIDS.
建立一种快速、高灵敏度的基于近红外荧光微球免疫层析法检测 P24 抗原的定量方法。
首先，我们制备了侧向流动检测试纸条，并使用荧光微球标记检测抗体。其次，我们优化了抗体标记条件。再次，我们优化了检测条件。然后，我们建立了工作曲线。最后，我们对建立的荧光微球免疫层析法进行了方法学评估。检测了 66 例临床样本，并将建立的荧光微球免疫层析法与定量 ELISA 法进行了比较。
根据工作曲线，该方法的检测限为 3.4pg/ml，检测范围为 3.4pg/ml 至 10ng/ml。平均批内回收率为 99.6%，变异系数（CV）为 5.4%-8.6%；平均批间回收率为 97.3%，CV 为 8.5%-11%。荧光微球免疫层析法的检测率高于 ELISA 法，与 ELISA 法具有良好的相关性。
基于近红外荧光微球免疫层析法的 P24 抗原定量检测方法具有检测快速、灵敏度高、检测范围广的优点；因此，它适用于艾滋病的早期临床诊断和连续监测。
