应用时间分辨荧光免疫分析技术检测SARS病毒抗原的初步研究

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (04): 429-431,434.

应用时间分辨荧光免疫分析技术检测SARS病毒抗原的初步研究

王蕾¹, 吴英松¹, 汤永平², 李明¹

1. 南方医科大学热带医学研究所, 广东, 广州, 510515;
2. 广州抗体技术研究中心, 广东, 广州, 510070

出版日期:2005-04-20 发布日期:2005-04-20
基金资助:
收稿日期:2004-10-23。
基金项目:军队非典防治紧急研究课题
作者简介:王蕾(1979-),女,2001年毕业于第一军医大学本科,现为在读硕士,电话:020-61640114-89133

Detection of SARS-associated coronavirus N protein by time-resolved fluoroimmunoassay

WANG Lei¹, WU Ying-song¹, TANG Yong-ping², LI Ming¹

1. 南方医科大学热带医学研究所, 广东, 广州, 510515;
2. 广州抗体技术研究中心, 广东, 广州, 510070

Online:2005-04-20 Published:2005-04-20

摘要/Abstract

摘要： 目的建立定量检测SARS冠状病毒N蛋白的时间分辨荧光免疫分析(TRFIA)。方法采用经SARS-CoVN蛋白和配对实验筛选的两株抗SARSN蛋白单克隆抗体,以双抗体夹心法为基础建立检测SARS-CoVN抗原的时间分辨荧光免疫分析技术,进行方法学的评价,并与相应ELISA试剂盒进行比较。结果该法的测量范围为(0.02~150)ng/ml,灵敏度为0.02ng/ml;批内、批间CV分别为(3.3~6.2)%和(5.3~9.6)%,与采用ELISA试剂盒检测灭活SARS-CoVN蛋白情况比较的结果一致。结论本研究建立的检测SARSN蛋白的时间分辨荧光免疫分析技术灵敏度高,特异性好,具有潜在的临床应用价值。

Abstract: Objective To develop a method for quantitative detection of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) N protein by timed-resolved fluoroimmunoassay (TRFIA). Methods Using a monoclonal antibody (mAb) against SARS-CoV N protein, screened by SARS-CoV N protein and matching experiment, a method for quantitative detection of SARS-CoV N protein by TRFIA was established on the basis of double sandwich enzyme-linked immunosorbent assay (ELISA) and evaluated against the ELISA kit. Result The measurement range of the assay was 0.02-150 ng/ml with a sensitivity of 0.02 ng/ml, the coefficient of variability within runs of 3.3%-6.2%, and coefficient of variability between days of 5.3%-9.6%. The results of detection were consistent between ELISA and TRFIA. Conclusion TRFIA is a new, sensitive and specific immunoassay for detecting SARS N protein with potential value in clinical applications.

中图分类号:

R392.33

王蕾¹, 吴英松¹, 汤永平², 李明¹. 应用时间分辨荧光免疫分析技术检测SARS病毒抗原的初步研究[J]. 南方医科大学学报, 2005, 25(04): 429-431,434.

WANG Lei¹, WU Ying-song¹, TANG Yong-ping², LI Ming¹. Detection of SARS-associated coronavirus N protein by time-resolved fluoroimmunoassay[J]. Journal of Southern Medical University, 2005, 25(04): 429-431,434.

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