Immunoreactivity characterisation of the three structural regions of the human coronavirus OC43 nucleocapsid protein by Western blot: implications for the diagnosis of coronavirus infection.
Immunoreactivity characterisation of the three structural regions of the human coronavirus OC43 nucleocapsid protein by Western blot: implications for the diagnosis of coronavirus infection.
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DOI：
10.1016/j.jviromet.2012.11.009
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发表时间：
2013-03
影响因子：
3.1
通讯作者：
Hou MH
中科院分区：
文献类型：
--
作者：
Liang FY;Lin LC;Ying TH;Yao CW;Tang TK;Chen YW;Hou MH
我们在超过90%的测试脐带血样本中检测到HCoV-OC 43 NP抗体。⑶ HCoV-OC 43 NP结构区的三种免疫反应模式。NP的中心连接区在人血清中呈高度免疫反应性。已有研究报道，原核表达的重组核衣壳蛋白（NP）是一种适用于冠状病毒感染流行病学筛查的试剂。本研究制备了可溶性重组人冠状病毒OC 43（HCoV-OC 43）NP，以检测β冠状病毒HCoV-OC 43 NP的抗原性。以纯化的重组NP为抗原，从兔血清中制备了HCoV-OC-43 NP特异性多克隆抗体，Western blot结果表明，该抗体与HCoV-OC-43 NP特异性反应，与其他人冠状病毒NP（包括SARS-CoV和HCoV-229 E）无交叉反应。同时检测了26例青年人、17例中老年呼吸道感染患者血清和15例脐带血标本。分别在96%、82%和93%的来自年轻人、呼吸道患者和脐带血样品的血清样品中观察到对HCoV-OC 43的NP的强反应性。为了鉴定NP的三个结构区域的免疫反应性，通过Western blot评估三个蛋白片段的抗原性，包括N-末端结构域（aa 1-173）、中心连接区（aa 174-300）和C-末端结构域（aa 301-448）。兔多克隆抗体表现出更大的免疫反应性的中央连接区和C-末端结构域比N-末端结构域。在人血清中观察到HCoV-OC 43 NP的三个结构区域的免疫反应性的三种不同模式，表明在人类HCoV-OC 43感染期间发生的免疫应答的可变性。NP的中心接头区域似乎是观察到的所有三种模式的最高免疫反应性区域。本研究的目的是提供深入了解HCoV感染的诊断工具的设计。
► We detected antibodies to HCoV-OC43 NP in over 90% of tested cord blood samples. ► Three patterns for the immunoreactivities of the structural regions of HCoV-OC43 NP. ► The central-linker region of the NP showed highly immunoreactive in human serum. Previous studies have reported that a prokaryotic-expressed recombinant nucleocapsid protein (NP) is a suitable reagent for the epidemiological screening of coronavirus infection. In this study, soluble recombinant human coronavirus OC43 (HCoV-OC43) NP was produced to examine the antigenicity of the HCoV-OC43 NP of betacoronavirus. Using the purified recombinant NP as an antigen, a polyclonal antibody from rabbit serum with specificity for HCoV-OC43 NP was generated; this antibody reacts specifically with HCoV-OC43 NP and does not cross-react with other human CoV NPs (including those of SARS-CoV and HCoV-229E) by Western blot. Sera from 26 young adults, 17 middle-aged and elderly patients with respiratory infection, and 15 cord blood samples were also tested. Strong reactivity to the NPs of HCoV-OC43 was observed in 96%, 82%, and 93% of the serum samples from the young adults, respiratory patients, and cord blood samples, respectively. To identify the immunoreactivities of the three structural regions of the NP that are recognised by the rabbit polyclonal antibody and human serum, the antigenicities of three protein fragments, including the N-terminal domain (aa 1-173), the central-linker region (aa 174-300), and the C-terminal domain (aa 301-448), were evaluated by Western blot. The rabbit polyclonal antibody demonstrated greater immunoreactivity to the central-linker region and the C-terminal domain than to the N-terminal domain. Three different patterns for the immunoreactivities of the three structural regions of HCoV-OC43 NP were observed in human serum, suggesting variability in the immune responses that occur during HCoV-OC43 infection in humans. The central-linker region of the NP appeared to be the most highly immunoreactive region for all three patterns observed. The goal of this study was to offer insight into the design of diagnostic tools for HCoV infection.
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影响因子：
1.8
作者：
Lee HK;Lee BH;Seok SH;Baek MW;Lee HY;Kim DJ;Na YR;Noh KJ;Park SH;Kumar DN;Kariwa H;Nakauchi M;Heo SJ;Park JH
通讯作者：
Park JH
影响因子：
3.1
作者：
Loa CC;Lin TL;Wu CC;Bryan TA;Hooper T;Schrader D
通讯作者：
Schrader D
影响因子：
3.1
作者：
Blanchard EG;Miao C;Haupt TE;Anderson LJ;Haynes LM
通讯作者：
Haynes LM
DOI：
10.1128/cdli.12.1.135-140.2005
发表时间：
2005-01-01
期刊：
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY
影响因子：
--
作者：
Di, B;Hao, W;Che, XY
通讯作者：
Che, XY
影响因子：
3.8
作者：
Nelson, GW;Stohlman, SA;Tahara, SM
通讯作者：
Tahara, SM