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. 2005 Sep 6;102(36):12891-6.
doi: 10.1073/pnas.0504666102. Epub 2005 Aug 23.

Cloning of a human parvovirus by molecular screening of respiratory tract samples

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Cloning of a human parvovirus by molecular screening of respiratory tract samples

Tobias Allander et al. Proc Natl Acad Sci U S A. .

Erratum in

  • Proc Natl Acad Sci U S A. 2005 Oct 25;102(43):15712

Abstract

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.

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Figures

Fig. 1.
Fig. 1.
Phylogenetic analysis: bootstrapped neighbor-joining tree based on full-length nucleotide sequences (Left) and ORF1 amino acid sequences (Right)of HBoV and the Parvovirinae. Bootstrap values are indicated at each branching point. Analysis of capsid gene nucleotide and amino acid sequences yielded highly similar results (data not shown). B19, erythrovirus B19; PTMPV, pig-tailed macaque parvovirus; LTMPV, long-tailed macaque parvovirus; RMPV, rhesus macaque parvovirus; ChPV, chipmunk parvovirus; AAV, adeno-associated virus; GPV, goose parvovirus; MDPV, Muscovy duck parvovirus; AMDV, Aleutian mink disease virus; PPV, porcine parvovirus; RPV-1a, rat parvovirus-1a; KRPV, Kilham rat parvovirus; MVM, minute virus of mice; CPV, canine parvovirus; FPLV, feline panleukopenia virus; MEV, mink enteritis virus.
Fig. 2.
Fig. 2.
Map of the HBoV genome. (A) Schematic map of isolate ST1 of HBoV showing the three ORFs as arrows: NS1, 1,920 nt (nucleotides 183-2102), 639 aa; NP-1, 660 nt (nucleotides 2340-2999), 219 aa; and VP1/VP2, 2,016 nt (nucleotides 2986-5001), 671 aa. (B) A map showing the location of the 26 nucleotide differences that were detected between two isolates of HBoV. The horizontal line represents the sequence of ST1, and each vertical line represents a nucleotide difference to ST2. In two cases where several differences were located close together, a longer vertical line representing four differences was used. The asterisks mark the three differences that resulted in a predicted amino acid change.

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