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The isotype of the mAb was analyzed by western blotting. Two clones named mAb11 and mAb12 were successfully established. Western blotting showed that the mAbs recognized both full-length nucleocapsid protein and cleaved nucleocapsid protein. These two mAbs also reacted with the nucleocapsid protein of other SARS-CoV strains, as determined by indirect ELISA. The mAbs were used to develop an ELISA. ELISA based on mAb11 and mAb12 showed higher sensitivity and specificity than commercial ELISA kits. The two mAbs and the ELISA could detect SARS-CoV antibody in serum samples obtained from SARS-CoV infected animals. Two mAbs against nucleocapsid protein were obtained, and were applied in the development of an ELISA to detect SARS-CoV antibody. The ELISA developed with mAb11 and mAb12 had higher sensitivity and specificity than the commercially available ELISA kits. 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