Viral genome sequencing by random priming methods

Autor: Zhang Xinsheng, Afonso Claudio, Feldblyum Jeremy, Sengamalay Naomi, DePasse Jay, Kuzmickas Ryan, Halpin Rebecca, Djikeng Appolinaire, Anderson Norman G, Ghedin Elodie, Spiro David J
Jazyk: angličtina
Rok vydání: 2008
Předmět:
Zdroj: BMC Genomics, Vol 9, Iss 1, p 5 (2008)
Druh dokumentu: article
ISSN: 1471-2164
DOI: 10.1186/1471-2164-9-5
Popis: Abstract Background Most emerging health threats are of zoonotic origin. For the overwhelming majority, their causative agents are RNA viruses which include but are not limited to HIV, Influenza, SARS, Ebola, Dengue, and Hantavirus. Of increasing importance therefore is a better understanding of global viral diversity to enable better surveillance and prediction of pandemic threats; this will require rapid and flexible methods for complete viral genome sequencing. Results We have adapted the SISPA methodology 123 to genome sequencing of RNA and DNA viruses. We have demonstrated the utility of the method on various types and sources of viruses, obtaining near complete genome sequence of viruses ranging in size from 3,000–15,000 kb with a median depth of coverage of 14.33. We used this technique to generate full viral genome sequence in the presence of host contaminants, using viral preparations from cell culture supernatant, allantoic fluid and fecal matter. Conclusion The method described is of great utility in generating whole genome assemblies for viruses with little or no available sequence information, viruses from greatly divergent families, previously uncharacterized viruses, or to more fully describe mixed viral infections.
Databáze: Directory of Open Access Journals