Immunoreactivity characterisation of the three structural regions of the human coronavirus OC43 nucleocapsid protein by Western blot: implications for the diagnosis of coronavirus infection.
Identifieur interne : 001158 ( PubMed/Checkpoint ); précédent : 001157; suivant : 001159Immunoreactivity characterisation of the three structural regions of the human coronavirus OC43 nucleocapsid protein by Western blot: implications for the diagnosis of coronavirus infection.
Auteurs : Fang-Ying Liang [Taïwan] ; Leng-Chieh Lin ; Tsung-Ho Ying ; Chen-Wen Yao ; Tswen-Kei Tang ; Yi-Wen Chen ; Ming-Hon HouSource :
- Journal of virological methods [ 1879-0984 ] ; 2013.
Descripteurs français
- KwdFr :
- Adolescent, Adulte, Adulte d'âge moyen, Animaux, Anticorps antiviraux (sang), Coronavirus humain OC43 (immunologie), Femelle, Humains, Infections à coronavirus (diagnostic), Jeune adulte, Mâle, Protéines nucléocapside (immunologie), Protéines recombinantes (immunologie), Sujet âgé, Sujet âgé de 80 ans ou plus, Technique de Western (), Techniques de laboratoire clinique ().
- MESH :
- diagnostic : Infections à coronavirus.
- immunologie : Coronavirus humain OC43, Protéines nucléocapside, Protéines recombinantes.
- sang : Anticorps antiviraux.
- Adolescent, Adulte, Adulte d'âge moyen, Animaux, Femelle, Humains, Jeune adulte, Mâle, Sujet âgé, Sujet âgé de 80 ans ou plus, Technique de Western, Techniques de laboratoire clinique.
English descriptors
- KwdEn :
- Adolescent, Adult, Aged, Aged, 80 and over, Animals, Antibodies, Viral (blood), Blotting, Western (methods), Clinical Laboratory Techniques (methods), Coronavirus Infections (diagnosis), Coronavirus OC43, Human (immunology), Female, Humans, Male, Middle Aged, Nucleocapsid Proteins (immunology), Recombinant Proteins (immunology), Young Adult.
- MESH :
- chemical , blood : Antibodies, Viral.
- diagnosis : Coronavirus Infections.
- immunology : Coronavirus OC43, Human, Nucleocapsid Proteins, Recombinant Proteins.
- methods : Blotting, Western, Clinical Laboratory Techniques.
- Adolescent, Adult, Aged, Aged, 80 and over, Animals, Female, Humans, Male, Middle Aged, Young Adult.
Abstract
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.
DOI: 10.1016/j.jviromet.2012.11.009
PubMed: 23174159
Affiliations:
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pubmed:23174159Le document en format XML
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<front><div type="abstract" xml:lang="en">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.</div>
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<Abstract><AbstractText>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.</AbstractText>
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<name sortKey="Lin, Leng Chieh" sort="Lin, Leng Chieh" uniqKey="Lin L" first="Leng-Chieh" last="Lin">Leng-Chieh Lin</name>
<name sortKey="Tang, Tswen Kei" sort="Tang, Tswen Kei" uniqKey="Tang T" first="Tswen-Kei" last="Tang">Tswen-Kei Tang</name>
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