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

doi:10.1016/j.jcv.2004.01.001

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

Research output: Contribution to journal › Article › Research › peer-review

32 Citations (Scopus)

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 language	English
Pages (from-to)	309-312
Number of pages	4
Journal	Journal of Clinical Virology
Volume	30
Issue number	4
DOIs	https://doi.org/10.1016/j.jcv.2004.01.001
State	Published - 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

Timani, K. A., Ye, L., Ye, L., Zhu, Y., Wu, Z., & Gong, Z. (2004). Cloning, sequencing, expression, and purification of SARS-associated coronavirus nucleocapsid protein for serodiagnosis of SARS. Journal of Clinical Virology, 30(4), 309-312. https://doi.org/10.1016/j.jcv.2004.01.001

Timani, Khalid A. ; Ye, Li ; Ye, Linbai ; Zhu, Ying ; Wu, Zhenghui ; Gong, Zuojiong. / Cloning, sequencing, expression, and purification of SARS-associated coronavirus nucleocapsid protein for serodiagnosis of SARS. In: Journal of Clinical Virology. 2004 ; Vol. 30, No. 4. pp. 309-312.

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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.",

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Timani, KA, Ye, L, Ye, L, Zhu, Y, Wu, Z & Gong, Z 2004, 'Cloning, sequencing, expression, and purification of SARS-associated coronavirus nucleocapsid protein for serodiagnosis of SARS', Journal of Clinical Virology, vol. 30, no. 4, pp. 309-312. https://doi.org/10.1016/j.jcv.2004.01.001

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 journal › Article › Research › peer-review

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