Isolation and Characterization of Viruses Related to the SARS Coronavirus from Animals in Southern China

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Science  10 Oct 2003:
Vol. 302, Issue 5643, pp. 276-278
DOI: 10.1126/science.1087139
  • Fig. 1.

    Detection of antibodies against recombinant nucleocapsid protein of SCoV in animal sera by Western blot assay. Recombinant nuleocapsid protein (NP, 49.6 kD) was used as an antigen to detect anti-SCoV antibodies in animal sera. Protein A-HRP was used as a secondary antibody, and reactive bands were visualized by the enhanced chemiluminesence Western blotting system. A serum sample from a convalescent SARS patient was used as a positive control. Blots reacted with animal (SZ2, SZ3, SZ11, SZ17, SZ7, SZ16, or SZ19) or human sera are indicated. Results from the neutralization test for SCoV-specific antibodies in these serum samples are also shown.

  • Fig. 2.

    Phylogenetic analysis of the nucleotide acid sequence of the spike gene of SCoV-like viruses. Nucleotide sequences of representative SCoV Sgenes (Sgene coding region 21477 to 25244, 3768 bp) were analyzed. The phylogenetic tree was constructed by the neighbor-joining method with bootstrap analysis (1000 replicates) using MEGA 2 (10). Number at the nodes indicates bootstrap values in percentage. The scale bar shows genetic distance estimated using Kimura's two-parameter substitution model (11). In addition to viruses sequenced in the present study, the other sequences used in the analysis could be found in GenBank with accession number: from AY304490 to AY304495, AY278741, AY278554, AY278491, AY274119, and AY278489.

  • Fig. 3.

    A 29-nt deletion in the human SCoV genome. (A) Genetic organization of SCoV-like viruses found in humans and animals. ORFs 1a and 1b, encoding the nonstructural polyproteins, and those encoding the S, E, M, and N structural proteins are indicated (green boxes). (B) Expanded view of the SCoV genomic sequence (27700 nt to 28200 nt, based on AY278554 numbering). ORFs for putative proteins and for N in human isolates are indicated as brown and green boxes, respectively (8). An extra 29-nt sequence is present downstream of the nucleotide of 27868 of the animal SCoV (based on AY278554 numbering). The presence of this 29-nt sequence in animals isolates results in fusing the ORFs 10 and 11 (top) into a new ORF (bottom; ORF10′, light blue box). (C) Protein sequence alignment of ORF10 and 11 from human isolates and ORF 10′ from animal isolates.

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    Isolation and Characterization of Viruses Related to the SARS Coronavirus from Animals in Southern China
    Y. Guan, B.J. Zheng, Y.Q. He, X. L. Liu, Z.X. Zhuang, C.L. Cheung, S.W. Luo, P.H. Li, L.J. Zhang, Y.J. Guan, K.M. Butt, K.L. Wong, K.W. Chan, W. Lim, K.F. Shortridge, K.Y. Yuen, J.S.M. Peiris, L.L.M. Poon

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