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Coronavirus Biology and Pathogenesis
Molecular Biology of Coronaviruses
Coronavirus Pathogenesis
Coronavirus Transmission and Persistence
Technology in SARS Discovery
Panel 1 Discussion
On the Front Lines
Approaches to Vaccines and Drug Development
Future Perspectives on Emerging Infections
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SARS in the Context of Emerging Infectious Threats SARS in the Context of Emerging Infectious Threats
Coronavirus Biology and Pathogenesis
Technology in SARS Discovery

Thomas Ksiazek, Centers for Disease Control and Prevention, Atlanta
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CDC involvement in SARS
• CDC Flu Branch became involved initially; Special Pathogens Branch became involved because of evidence of person-to-person transmission.
• Special Pathogens focused on obtaining diagnostic specimens in early days, working to identify specific diagnostic tests, the pathological process, possible agents, and tissues to focus on.
• Presence of giant cells in lung and diffuse alveolar damage suggested a viral etiology.

Molecular Techniques
• Vero E6 cells were inoculated.
• Electron microscopy, negative stain, thin-section morphology, and morphogenesis resembled coronaviruses.
• Dean Erdman's group designed primers that can be used as a diagnostic technique on clinical materials.
• Sequence analysis suggests SARS virus is the first in a new group of coronaviruses.
• Cofactors still may be involved in causing SARS, but coronavirus is the etiologic agent.
• Antigens could not be found on giant cells from patients, but antigens usually do not persist for long in many respiratory infections.

Diagnostic Techniques
• Vero E6 cells can be stained in an indirect fluorescent antibody (IFA) with convalescent patient serum, showing viral antigens present.
• An ELISA test was developed using antigens from infected cells.
• Western blot on convalescent patient materials show N protein predominates and is goal for recombinant protein synthesis.
• ELISA and IFA can be used on convalescent and acute sera from patients to show conversion from negative to positive.

SARS in Cultures and Tissues
• Virus sequences from various sites is essentially identical.
• The disease is reproduced in cynomolgus monkeys.
• Virus is identified in respiratory secretions and tissues using a number of techniques.
• Patients, including those who succumb, have neutralizing antibodies.

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