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Recovery of human metapneumovirus genetic lineages a and B from cloned cDNA. | LitMetric

AI Article Synopsis

  • Human metapneumovirus (hMPV) is a recently identified virus that mainly affects respiratory health in young children, the elderly, and people with weakened immune systems.
  • The study details the sequencing of leader and trailer regions of two specific hMPV isolates, showcasing their genetic differences and providing insight into the virus's biology.
  • Researchers successfully developed a reverse genetics system that allows for the creation of recombinant viruses, which can aid in further research and understanding of hMPV behavior and treatment options.

Article Abstract

Human metapneumovirus (hMPV) is a newly discovered pathogen associated with respiratory tract illness, primarily in young children, immunocompromised individuals, and the elderly. The genomic sequence of the prototype hMPV isolate NL/1/00 without the terminal leader and trailer sequences has been reported previously. Here we describe the leader and trailer sequences of two hMPV isolates, NL/1/00 and NL/1/99, representing the two main genetic lineages of hMPV. Minigenome constructs in which the green fluorescent protein or chloramphenicol acetyltransferase genes are flanked by the viral genomic ends derived from both hMPV lineages and transcribed using a T7 RNA polymerase promoter-terminator cassette were generated. Cotransfection of minigenome constructs with plasmids expressing the polymerase complex components L, P, N, and M2.1 in 293T or baby hamster kidney cells resulted in expression of the reporter genes. When the minigenome was replaced by a sense or antisense full-length cDNA copy of the NL/1/00 or NL/1/99 viral genomes, recombinant virus was recovered from transfected cells. Viral titers up to 10(7.2) and 10(5.7) 50% tissue culture infective dose/ml were achieved with the sense and antisense plasmids, respectively. The recombinant viruses replicated with kinetics similar to those of the parental viruses in Vero cells. This reverse genetics system provides an important new tool for applied and fundamental research.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC446134PMC
http://dx.doi.org/10.1128/JVI.78.15.8264-8270.2004DOI Listing

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