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KMID : 0545120080180101717
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Journal of Microbiology and Biotechnology
2008 Volume.18 No. 10 p.1717 ~ p.1721
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Detection of Antibodies Against SARS-Coronavirus Using Recombinant Truncated Nucleocapsid Proteins by ELISA
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Lee Hyun-Kyoung
Lee Byoung-Hee
Dutta Noton Kumar
Seok Seung-Hyeok
Baek Min-Won
Lee Hui-Young
Kim Dong-Jae
Na Yi-Rang
Noh Kyoung-Jin
Park Sung-Hoon
Kariwa Hiroaki
Nakauchi Mina
Mai Le Quynh
Heo Suk-Jin
Park Jae-Hak
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Abstract
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Severe acute respiratory syndrome (SARS) is a lifethreatening emerging respiratory disease caused by the coronavirus, SARS-CoV. The nucleocapsid (N) protein of SARS-CoV is highly antigenic and may be a suitable candidate for diagnostic applications. We constructed truncated recombinant N proteins (N1 [1-422 aa], N2 [1- 109 aa], and N3 [110-422 aa]) and determined their antigenicity by Western blotting using convalescent SARS serum. The recombinants containing N1 and N3 reacted with convalescent SARS serum in Western blotting. However, the recombinant with N2 did not. In ELISA using N1 or N3 as the antigens, positive results were observed in 10 of 10 (100%) SARS-CoV-positive human sera. None of 50 healthy sera gave positive results in either assay. These data indicate that the ELISA using N1 or N3 has high sensitivity and specificity. These results suggest that the middle or C-terminal region of the SARS N protein is important for eliciting antibodies against SARS-CoV during the immune response, and ELISA reactions using N1 or N3 may be a valuable tool for SARS diagnosis.
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KEYWORD
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ELISA, SARS, truncated recombinant nucleocapsid proteins
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