Isolation Of Human Metapneumovirus, Expression Of Fusion Protein And Immunoinformatic Analysis Of Major Proteins

PARTâ… First Isolation and Characterization of Human Metapneumovirus in Mainland ChinaObjective: To isolate and characterize the newly discoveried human metapneumovirus in mainland China and then elucidate the preliminary molecular epidemiology about it.Methods: The patients hospitalized for RTIs in the Children's Hospital, Chongqing University of Medical sciences were enrolled in the present study. Nasopharyngeal aspirates (NPAs) were collected for common respiratory viruses screening by direct immunofluorescence assay and for inoculation onto Vero-E6 cells and LLC-MK2 cells for hMPV isolation. Cultures were incubated in viral growth medium with trypsin and observed for the development of cytopathic effect (CPE) for 3 weeks. Positive ones were confirmed by subsequent reverse transcription polymerase chain reaction targeting N, F genes of hMPV. The amplified F fragments sequence of the isolates were determined and then used for phylogenetic tree construction by neighbor-joining algorithm and maximum-parsimony algorithm in the Phylip package and multi-alignment by CLUSTAL X software. The clinic findings of the children with hMPV infection were collected and analyzed.Results: Of the collected 86 NPAs, six ones showed CPE. At different stages post infection, increased granules or detachment from cell monolayer but no obvisious syncytium were observed. Successful isolation...