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
Department of Laboratory Animal Medicine, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea.
J Microbiol Biotechnol. 2008 Oct;18(10):1717-21.
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.
严重急性呼吸综合征（SARS）是一种由冠状病毒SARS-CoV引起的危及生命的新发呼吸道疾病。SARS-CoV的核衣壳（N）蛋白具有高度抗原性，可能是诊断应用的合适候选物。我们构建了截短的重组N蛋白（N1 [1-422个氨基酸]、N2 [1-109个氨基酸]和N3 [110-422个氨基酸]），并使用恢复期SARS血清通过蛋白质印迹法测定它们的抗原性。含有N1和N3的重组体在蛋白质印迹法中与恢复期SARS血清发生反应。然而，含有N2的重组体则没有。在以N1或N3作为抗原的酶联免疫吸附测定（ELISA）中，10份SARS-CoV阳性人血清中的10份（100%）检测结果呈阳性。50份健康血清在这两种检测中均未得出阳性结果。这些数据表明，使用N1或N3的ELISA具有高灵敏度和特异性。这些结果表明，SARS N蛋白的中部或C末端区域对于在免疫反应期间引发针对SARS-CoV的抗体很重要，并且使用N1或N3的ELISA反应可能是SARS诊断的一种有价值的工具。
