Cloning, sequencing, expression, and purification of SARS-associated coronavirus nucleocapsid protein for serodiagnosis of SARS

Khalid A. Timani, Li Ye, Linbai Ye, Ying Zhu, Zhenghui Wu, Zuojiong Gong

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Abstract

A novel coronavirus has been associated with a worldwide outbreak of atypical pneumonia referred to as Severe Acute Respiratory Syndrome (SARS-CoV). SARS-CoV nucleocapsid (N) protein has been cloned sequenced and expressed in Escherichia coli strain. Purified N protein was used to measure the SARS-CoV specific IgG antibodies from 16 SARS-CoV infected patients' sera and from 131 control subjects using ELISA assay. Specific antibody responses to the purified recombinant N protein after 10, 20, and 30 days of disease onset were observed in 13 of 16 (81.3%), 16 of 16 (100%) and 16 of 16 (100%) SARS patients sera, respectively. Comparison of detection results with a commercially available diagnostic kit coated with a mixture of SARS-CoV viral proteins showed 9 of 16 (56.3%), 13 of 16 (81.3%), and 15 of 16 (93.7%) positive responses, respectively. None of 131 control sera gave positive reaction in either assay. This data suggests that the N protein of SARS-CoV is immunodominant and this ELISA based test assay for detecting the SARS-CoV N antigen may hold a significant value for SARS diagnosis.

Original languageEnglish
Pages (from-to)309-312
Number of pages4
JournalJournal of Clinical Virology
Volume30
Issue number4
DOIs
StatePublished - 1 Aug 2004

Fingerprint

SARS Virus
Serologic Tests
Organism Cloning
Serum
Enzyme-Linked Immunosorbent Assay
Nucleocapsid Proteins
Severe Acute Respiratory Syndrome
Coronavirus
Viral Proteins
Recombinant Proteins
Antibody Formation
Disease Outbreaks
Pneumonia
Immunoglobulin G
Coronavirus nucleocapsid protein
Escherichia coli
Antigens
Antibodies
Proteins

Keywords

  • ELISA
  • Expression and purification
  • Nucleocapsid protein
  • SARS-CoV

Cite this

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title = "Cloning, sequencing, expression, and purification of SARS-associated coronavirus nucleocapsid protein for serodiagnosis of SARS",
abstract = "A novel coronavirus has been associated with a worldwide outbreak of atypical pneumonia referred to as Severe Acute Respiratory Syndrome (SARS-CoV). SARS-CoV nucleocapsid (N) protein has been cloned sequenced and expressed in Escherichia coli strain. Purified N protein was used to measure the SARS-CoV specific IgG antibodies from 16 SARS-CoV infected patients' sera and from 131 control subjects using ELISA assay. Specific antibody responses to the purified recombinant N protein after 10, 20, and 30 days of disease onset were observed in 13 of 16 (81.3{\%}), 16 of 16 (100{\%}) and 16 of 16 (100{\%}) SARS patients sera, respectively. Comparison of detection results with a commercially available diagnostic kit coated with a mixture of SARS-CoV viral proteins showed 9 of 16 (56.3{\%}), 13 of 16 (81.3{\%}), and 15 of 16 (93.7{\%}) positive responses, respectively. None of 131 control sera gave positive reaction in either assay. This data suggests that the N protein of SARS-CoV is immunodominant and this ELISA based test assay for detecting the SARS-CoV N antigen may hold a significant value for SARS diagnosis.",
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Cloning, sequencing, expression, and purification of SARS-associated coronavirus nucleocapsid protein for serodiagnosis of SARS. / Timani, Khalid A.; Ye, Li; Ye, Linbai; Zhu, Ying; Wu, Zhenghui; Gong, Zuojiong.

In: Journal of Clinical Virology, Vol. 30, No. 4, 01.08.2004, p. 309-312.

Research output: Contribution to journalArticleResearchpeer-review

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