1. 新乡医学院医学检验学院
2. ,广东,广州,510642
3. 河南省生物技术研究中心
4. 郑州市疾病预防控制中心
5. 河南省疾病预防控制中心
6. 河南中医药大学第二临床医学院
7. 河南省职工医院
8. 郑州职业技术学院
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卢瑶, 牛卫东, 徐瑾, 等. 截短新型冠状病毒N蛋白在荧光层析法上的应用[J]. 分析测试学报, 2021,40(5):705-711.
Application of Truncated SARS-CoV-2 N Protein in Fluorescence Immunochromatography[J]. 2021,40(5):705-711.
基于截短新型冠状病毒(Severe Acute Respiratory Syndrome Coronavirus 2,SARS-CoV-2)N蛋白和全长N蛋白,采用荧光免疫层析技术,建立了一种高灵敏度快速评价新型冠状病毒肺炎患者体内SARS-CoV-2抗体水平的方法。以荧光微球为标记物制备试纸条,进行阴性和阳性检测并选出较优抗原配对组合。初步评价了其精密度与稳定性,并与已上市的检测方法进行对比。根据阴性和阳性检测结果优选截短N蛋白标记,全长N蛋白包被组合,所建立的方法可以在 15 min内完成SARS-CoV-2抗体水平的检测,能够有效区分阴阳性,批内精密度CV为3.48%~10.05%,批间精密度CV为4.77%~11.73%。试纸条在37 ℃条件下能够稳定保存21 d,与胶体金检测法一致性较好,总符合率为96.13%,阳性符合率为90.63%,阴性符合率为97.56%。该研究基于截短N蛋白初步建立的荧光免疫层析检测方法能有效评价患者体内的SARS-CoV-2抗体水平,具有良好的精密度和稳定性,便于在基层医疗单位推广,有较好的应用前景。
A time-resolved immunofluorescence chromatography was established for the rapid and sensitive evaluation of SARS-CoV-2 antibody level in patients suffering Corona Virus Disease 2019,with a truncated severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) N protein and a full-length SARS-CoV-2 N protein.The test strip cards were prepared with fluorescent microspheres as markers,which were assembled for negative and positive detection to pick out the superior antigen pairing combination.Meanwhile,the precision and stability of the optimized test strip card were further evaluated for preliminary performance,and compared with those of the marketed SARS-CoV-2 antibody detection method on this basis.According to the negative and positive detection results,preferably,the truncated N protein was labeled and the full-length N protein was coated.This method could complete the detection of SARS-CoV-2 antibody level within 15 minutes,and could effectively distinguish the negative and positive results.The intra-assay precision CV was between 3.48% and 10.05%,and the inter-assay precision CV was between 4.77% and 11.73%.The strips could be stably stored at 37 ℃ for 21 days.The detection results by this method were well correlated with those by colloidal gold assay,with a total coincidence rate of 96.13%,a positive coincidence rate of 90.63% and a negative coincidence rate of 97.56%,respectively.The fluorescence immunochromatographic assay preliminarily established based on truncated N protein could effectively evaluate the SARS-CoV-2 antibody level in patients with good precision and stability,which was convenient for popularization in primary care units and had a good application prospect.
新型冠状病毒新型冠状病毒肺炎N蛋白荧光免疫层析检测
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)Corona Virus Disease 2019(COVID-19)N proteinfluorescence immunochromatographydetection
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