A New Method for Ultra-sensitive P24 Antigen Assay Based on Near-infrared Fluorescent Microsphere Immunochromatography
A New Method for Ultra-sensitive P24 Antigen Assay Based on Near-infrared Fluorescent Microsphere Immunochromatography
复制标题
基于近红外荧光微球免疫层析的超灵敏P24抗原测定新方法
DOI：
10.3967/bes2020.024
复制
发表时间：
2020-03-01
影响因子：
3.5
通讯作者：
Cao Yu Xi
中科院分区：
文献类型：
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作者：
Wang Qi;Hou Mei Ling;Cao Yu Xi
目的建立一种快速、高灵敏度的近红外荧光微球免疫层析定量检测P24抗原的方法。其次，我们优化了抗体标记的条件。第三，优化了检测条件。第四，我们创造了一条工作曲线。第五，对建立的荧光微球免疫层析方法进行了方法学评价。结果建立的荧光微球免疫层析法的检测限为3.4 pg/mL，检测范围为3.4 ~ 10 ng/mL。批内平均回收率为99.6%，变异系数（CV）为5.4%~ 8.6%;批间平均回收率为97.3%，CV为8.5%~ 11%。结论基于近红外荧光微球免疫层析的P24抗原定量检测方法具有快速、灵敏度高、检测范围广等优点，适用于艾滋病的早期临床诊断和持续监测。
Objective To develop a rapid, highly sensitive quantitative method for detecting P24 antigen based on near-infrared fluorescent microsphere immunochromatography.Methods 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.Results 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.Conclusion 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.