SARS- and Other Coronaviruses: Laboratory Protocols

The year 2003 was the year when the name “coronavirus” went around the world, somewhat further than the virus that sparked panic: severe acute respira­tory syndrome coronavirus (SARS-CoV). It was spread rapidly by international, indeed transcontinental, travelers from its epicenter in China. The high mortality rate, around 10% among clinical cases, and the particularly high price paid by health care workers, spread fear globally. Public health facilities were stretched to the limit, and the effect on local economies was immense. Never before had a coronavirus made such an impact on the lives of the human population.

SARS enhanced interest in coronaviruses generally, including the hunt not only for SARS-CoV-like viruses in wild animals, but also for unrelated coro- naviruses in wild and semi-domesticated animals. Prior to SARS we knew of only a dozen or so coronavirus species but it was always likely that there were many more. Coronavirologists knew, from decades of experience, that coron- aviruses could be devastating, causing mortality, especially among young, and high economic loss among domestic animals. Moreover, they knew that a given coronavirus species was not limited to replication in one host species. Thus the potential existed for the spread of novel coronaviruses from wild animals to domestic animals and to man. It is with this potent threat in mind that I have included no fewer than seven chapters dealing with the detection and discovery of coronaviruses by nucleic acid approaches, with antibody-based approaches being described in three other chapters.

Although one can detect a virus these days without having to grow it in the laboratory, there is always a need to do this at some point, whether it be for the development of diagnostics and vaccines or for the study of pathogene­sis, pathogenicity, variation, and other virological properties. Therefore, I have included several chapters on virus isolation and propagation.

For some purposes, e.g., structural studies (Section 3) and vaccine devel­opment, it is necessary to rigorously purify a virus after propagating it, an art that fewer virologists practice these days. Hence, two chapters contain detailed procedures for this. For other structural, and functional, studies and for raising antibodies for subsequent analytical use, it is sufficient, and, in the case of nonstructural proteins, essential, to express viral proteins individually. Several chapters address this task, including one that deals with crystallization of non- structural proteins.

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