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

TLDR

Identifying new virus species is crucial for infectious disease research but remains technically challenging. The authors propose initiating a systematic exploration of the human virome. They developed a large‑scale, culture‑independent screening system that depletes host DNA, performs random PCR amplification, and uses high‑throughput sequencing and bioinformatics, applied to pooled respiratory tract samples. The method first identified seven human viruses—including an uncharacterized coronavirus and a novel parvovirus—without specific reagents; the parvovirus, later named human bocavirus, was found in 17 additional pediatric patients with lower respiratory tract infections, demonstrating the approach’s utility for detecting unknown viruses.

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.