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Recovery of a Paramyxovirus, the Human Metapneumovirus, from Cloned cDNA

  • B. G. van den HoogenEmail author
  • R. A. M. Fouchier
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1602)

Abstract

Human metapneumovirus (HMPV), a single-stranded negative-sense RNA virus belonging to the family Paramyxoviridae, is associated with respiratory tract illness, primarily in young children and persons with underlying disease. Based on genetic and antigenic variation, HMPV strains are classified into two serotypes, with isolates NL/1/00 and NL/1/99 as prototypes for serotypes A and B, respectively. The development of plasmid-based reverse genetics systems for both serotypes has resulted in developments of a wide range of vaccine candidates against HMPV infection. The approach to virus rescue of HMPV is similar to that used for other paramyxoviruses, starting with mini-replicon assays for optimizations of the rescue protocols and subsequent replacement of the mini genome with a plasmid expressing the cDNA of the full-length viral RNA genome. Here, we provide detailed information on the reverse genetics systems for HMPV.

Key words

Human metapneumovirus HMPV Single-stranded negative RNA virus Reverse genetics Virus rescue 

Notes

Acknowledgments

We like to thank M. de Graaf and S. Herfst for their valuable contribution to the research field of human metapneumovirus, through the development of the rescue genetics systems.

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© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Department of ViroscienceErasmus MC, University Medical Center RotterdamRotterdamThe Netherlands

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