Novel rapid immunochromatographic test based on an enzyme immunoassay for detecting nucleocapsid antigen in SARS-associated coronavirus
Novel rapid immunochromatographic test based on an enzyme immunoassay for detecting nucleocapsid antigen in SARS-associated coronavirus
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DOI：
10.1002/jcla.20070
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发表时间：
2005-01-01
影响因子：
2.7
通讯作者：
Okada, M
中科院分区：
文献类型：
--
作者：
Kogaki, H;Uchida, Y;Okada, M
一种新型严重急性呼吸综合征相关冠状病毒（SARS-CoV）被发现。从疑似SARS患者标本中检测SARS冠状病毒抗原和抗体对预防感染具有重要作用。本研究利用重组N-Ag单克隆抗体（MoAbs）免疫小鼠，建立了一种基于夹心型酶免疫分析法（EIA）的新型快速免疫层析（RICT）检测SARS-CoV天然核衣壳抗原（N-Ag）的方法。RICT是一种用于呼吸道吸入物和血清标本的定性分析方法。使用此分析，样品应用15分钟后，可以通过设备上蓝线的出现来主观判断阳性结果。结合多对MoAbs的RICT对SARS-CoV重组N-Ag和病毒N-Ag的检测均具有较高的敏感性。rSN122和rSN21-2分别是固定化抗体和酶标记的最佳MoAbs。在分析灵敏度方面，重组N-Ag的检测浓度为31 pg/mL， SARS-CoV的检测浓度为1.99 × 10(2) TCID50/mL。当使用150份人血清和50份鼻咽吸痰（NSPs）时，RICT的特异性为100%。基于rSN122/rSN21-2对EIA的RICT是一种敏感、特异、可靠的快速检测方法，可用于热处理或Triton X-100处理的SARS-CoV中n -银的检测。
A novel severe acute respiratory syndrome-associated coronavirus (SARS-CoV) has been discovered. The detection of both antigens and antibodies in SARS-CoV from human specimens with suspected SARS plays an important role in preventing infection. We developed a novel rapid immunochromatographic test (RICT) based on the sandwich format enzyme immunoassay (EIA) with an all-in-one device for detecting the native nucleocapsid antigen (N-Ag) of SARS-CoV using monoclonal antibodies (MoAbs), which we produced by immunizing recombinant N-Ag to mice. RICT is a qualitative assay for respiratory aspirates and serum specimens. With this assay, a positive result can be judged subjectively by the appearance of a blue line on the device 15 min after the sample is applied. RICT with several pairs of MoAbs showed a high sensitivity for the detection of recombinant N-Ag as well as viral N-Ag of SARS-CoV. rSN122 and rSN21-2 were the best MoAbs for immobilized antibody and enzyme labeling, respectively. With regard to analytical sensitivity, RICT detected N-Ag at 31 pg/mL for recombinant N-Ag, and at 1.99x10(2) TCID50/mL for SARS-CoV. The specificity of RICT was 100% when 150 human sera and 50 nasopharyngeal aspirates (NSPs) were used. RICT based on an EIA using the rSN122/rSN21-2 pair is a sensitive, specific, and reliable rapid assay for detecting N-Ag in SARS-CoV treated with either heat or Triton X-100.