Cloning of a Human Parvovirus by Molecular Screening of Respiratory Tract Samples

The identification of new virus species is a key issue for the study of infectious disease but is technically very difficult. We developed a system for large-scale molecular virus screening of clinical samples based on host DNA depletion, random PCR amplification, large-scale sequencing, and bioinfo...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2005-09, Vol.102 (36), p.12891-12896
Main Authors: Allander, Tobias, Tammi, Martti T., Eriksson, Margareta, Bjerkner, Annelie, Tiveljung-Lindell, Annika, Andersson, Björn, Palese, Peter
Format: Article
Language:English
Subjects:
Biological Sciences
Bocavirus
Child, Preschool
Cloning, Molecular
Coronavirus
DNA
Etiology
Female
Genome, Viral
Humans
Incidence
Infant
Infections
Male
Medicin och hälsovetenskap
Molecular Sequence Data
Nasopharynx - virology
Nucleotides
Parvovirus
Parvovirus - genetics
Parvovirus - isolation & purification
Parvovirus - physiology
Phylogeny
Polymerase chain reaction
Respiratory Tract Diseases - virology
Sequencing
Viruses
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Summary:The identification of new virus species is a key issue for the study of infectious disease but is technically very difficult. We developed a system for large-scale molecular virus screening of clinical samples based on host DNA depletion, random PCR amplification, large-scale sequencing, and bioinformatics. The technology was applied to pooled human respiratory tract samples. The first experiments detected seven human virus species without the use of any specific reagent. Among the detected viruses were one coronavirus and one parvovirus, both of which were at that time uncharacterized. The parvovirus, provisionally named human bocavirus, was in a retrospective clinical study detected in 17 additional patients and associated with lower respiratory tract infections in children. The molecular virus screening procedure provides a general culture-independent solution to the problem of detecting unknown virus species in single or pooled samples. We suggest that a systematic exploration of the viruses that infect humans, "the human virome," can be initiated.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0504666102