Abstract:
Objective To analyze the gene sequence of Hantavirus (HV) isolates in Fujian province in 2019 at molecular level and provide experimental evidence for the prevention and control hemorrhagic fever with renal syndrome (HFRS).
Methods The serum samples of patients with acute HFRS and rat lung specimens were collected from 2018 to 2020. HV IgM IgG antibody in the serum sample were detected by colloidal gold, HV antigen in rat lung specimens were detected by immunofluorescence assay test. Positive samples were inoculated with E6 cell line to isolate virus. The supernatants of the cultures of specimens were detected by reverse transcription-polymerase chain reaction and the cells were detected by immunofluorescence assay test. The genome of HV isolates were sequenced using high-throughput sequencing technique. Finally, the sequence alignment and phylogenetic analysis were conducted with software MEGA 11.0.
Results A total of 879 rodents were captured, and the positive rate was 2.84%. Two strains of Hantavirus were isolated from Apodemus agrarius. Genome sequence analysis showed Hantaan virus (HTNV). Phylogenetic analysis using the maximum likelihood method showed that the isolates belonged an distinct lineage and formed a clade with strains isolated in Zhejiang (longquanAA-10-101, Z251 and LongquanMF-08-92), showing 95.81% nt sequence identity. Compared with vaccine strain Z10, the amino variations of Thr43Ala and Lys95Met were observed at nucleoproteins of isolates. Compared with HTNV international reference strain 76-118 and vaccine strain 84FLi, the isolates lacked glycoylation site at N766.
Conclusion Hantavirus isolates from Fujian had unique genetic characteristics and exhibited a close evolutionary relationship with strains isolated in Zhejiang which might derive from co-evolving ancestors. The effect of amino variations in envelope glycoproteins and nucleocapsid proteins of Fujian isolates need further research.
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