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

Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology
Khalid Amine TimaniZuojiong Gong

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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Citations

Oct 16, 2007·Clinical Microbiology Reviews·Vincent C C ChengKwok Yung Yuen
Feb 5, 2008·Biochemical and Biophysical Research Communications·Yingchun ZengZhenghui Wu
Aug 2, 2005·Journal of Biochemistry and Molecular Biology·Xiaonan FangZhenghui Wu
May 6, 2020·Viruses·Maddalena DiluccaAthanasia Pavlopoulou
Sep 7, 2005·Acta Biochimica Et Biophysica Sinica·Qing-Jiao LiaoZheng-Hui Wu
Dec 17, 2004·Journal of Medical Virology·Paul K S ChanPak Leong Lim
Feb 2, 2021·Measurement : Journal of the International Measurement Confederation·Madeshwari EzhilanNoel Nesakumar

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Datasets Mentioned

BETA
AY365036

Methods Mentioned

BETA
PCR
electrophoresis
ELISA

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