Department of Pediatrics, Fukushima Medical University.
Fukushima J Med Sci. 2022 Dec 21;68(3):153-159. doi: 10.5387/fms.2022-15. Epub 2022 Aug 31.
This study investigated whether quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), using specific probes composed of locked nucleic acids (LNA/qRT-PCR), designed to evaluate H1N1 pdm09 H275Y, H3N2 E119V and R292K variant populations, could replace a neuraminidase (NA) inhibition assay to determine the 50% inhibitory concentration (IC) of NA activity.For H1N1 pdm09, when the H275Y variant RNA load was 50% or 70% and the infective H275Y variant load was 40% or 70%, the IC were >10- and 100-fold higher, respectively, than that of the wild-type (WT) strain. For H3N2, when the E119V RNA load and infective E119V variant load were >90% and >60%, respectively, the IC of the mixed sample was >10-fold higher than that of the WT strain. The variant-mixed samples with a 70% or 80% R292K variant RNA load and a 60% or 70% infective R292K variant load exhibited >10- and 100-fold decreased susceptibility, respectively, compared with that of the WT. A positive correlation between the variant RNA load and infective variant load populations was observed.The LNA/qRT-PCR method can be used to improve the treatment and management of patients during antiviral therapy for influenza virus infection.
本研究旨在探讨使用针对 H1N1 pdm09 H275Y、H3N2 E119V 和 R292K 变异株设计的、包含锁核酸（LNA）的实时荧光定量逆转录聚合酶链反应（qRT-PCR）（LNA/qRT-PCR），是否可以替代神经氨酸酶（NA）抑制测定法来确定 NA 活性的 50%抑制浓度（IC）。对于 H1N1 pdm09，当 H275Y 变异 RNA 载量为 50%或 70%，感染性 H275Y 变异载量为 40%或 70%时，IC 值分别比野生型（WT）株高 10 倍和 100 倍。对于 H3N2，当 E119V RNA 载量和感染性 E119V 变异载量分别大于 90%和 60%时，混合样本的 IC 值比 WT 株高 10 倍以上。当 70%或 80%的 R292K 变异 RNA 载量和 60%或 70%的感染性 R292K 变异载量时，变异混合样本的敏感性分别比 WT 株降低 10 倍和 100 倍。观察到变异 RNA 载量与感染性变异株群体之间存在正相关。LNA/qRT-PCR 方法可用于改善流感病毒感染患者在抗病毒治疗期间的治疗和管理。
