Abed Yacine, Goyette Nathalie, Boivin Guy
Research Center in Infectious Diseases of the CHUQ-CHUL, Québec City, Canada.
Antivir Ther. 2004 Aug;9(4):577-81.
A system of reverse genetics was used to generate influenza A/H1N1 viruses harbouring neuraminidase (NA) mutations previously associated with resistance to NA inhibitors in various viral subtypes. The His274Tyr and Glu119Gln mutants were rescued whereas the Arg292Lys and Glu1l9 --> Gly, Val, Ala or Asp mutants could not be generated. In NA inhibition assays, the His274Tyr mutant was resistant to oseltamivir (430-fold over wild-type) and BCX-1812 (50-fold) but was sensitive to zanamivir. A similar trend was seen when the mutant was evaluated by plaque reduction assay (PRA). The Glu119Gln mutant expressed a low level of resistance to oseltamivir (nine-fold) and zanamivir (fourfold) in NA inhibition assay but was only marginally resistant to oseltamivir (fourfold) in PRA. The replication capacity of both mutants, in particular that of the His274Tyr virus, was impaired when compared with the wild-type virus in vitro.
利用反向遗传学系统生成了携带神经氨酸酶（NA）突变的甲型H1N1流感病毒，这些突变先前与多种病毒亚型中对NA抑制剂的耐药性有关。His274Tyr和Glu119Gln突变体得以拯救，而Arg292Lys和Glu119→Gly、Val、Ala或Asp突变体则无法产生。在NA抑制试验中，His274Tyr突变体对奥司他韦（比野生型高430倍）和BCX-1812（50倍）耐药，但对扎那米韦敏感。通过蚀斑减少试验（PRA）评估该突变体时也观察到类似趋势。在NA抑制试验中，Glu119Gln突变体对奥司他韦（9倍）和扎那米韦（4倍）表现出低水平耐药，但在PRA中仅对奥司他韦有轻微耐药（4倍）。与野生型病毒相比，两种突变体，尤其是His274Tyr病毒的体外复制能力均受损。
