Zeberezhnyĭ A D, Grebennikova T V, Vorkunova G K, Yuzhakov A G, Kostina L V, Norkina S N, Aliper T I, Nepoklonov E A, Lvov D K
Vopr Virusol. 2014;59(6):23-7.
Reverse genetics was applied to engineering of the reassortantvaccine candidate strain against highly pathogenic avian influenza viruses (HPAIVs) of the H5 subtype. The new strain recPR8-H5N1 contains the HA gene from the Russian HPAIV A/Kurgan/05/2005 (H5N1), the NA and internal genes from A/Puerto Rico/8/34 (H1N1). The strain recPR8-H5N1 demonstrated the antigenic specificity (H5), high proliferation rate in 12 days chicken embryos, and was lethal for the embryos in 36 hours. An inactivated emulsified vaccine based on the strain recPR8-H5N1 elicited high antibody titers and protected 6-week-old chickens from lethal challenge with the HPAIV A/Kurgan/05/2005 (H5N1) on day 21 after single immunization. Infection of non-vaccinated birds with the strain recPR8-H5N1 did not cause any pathology, and the virus was not detected using PCR in blood and cloacal swabs on day 7 p.i. Specific weak seroconversion caused by infection with the strain recPR8-H5N1 was detected on day 14 p.i. As a result, a new influenza virus strain was obtained with modified properties.
反向遗传学被应用于针对H5亚型高致病性禽流感病毒(HPAIVs)的重组候选疫苗株的工程改造。新毒株recPR8-H5N1包含来自俄罗斯HPAIV A/Kurgan/05/2005(H5N1)的血凝素(HA)基因、来自A/波多黎各/8/34(H1N1)的神经氨酸酶(NA)基因和内部基因。recPR8-H5N1毒株表现出抗原特异性(H5),在12日龄鸡胚中增殖率高,且在36小时内对鸡胚致死。基于recPR8-H5N1毒株的灭活乳化疫苗可诱导产生高抗体滴度,并在单次免疫后第21天保护6周龄鸡免受HPAIV A/Kurgan/05/2005(H5N1)致死性攻击。用recPR8-H5N1毒株感染未接种疫苗的禽类未引起任何病变,且在感染后第7天,血液和泄殖腔拭子中未通过聚合酶链反应(PCR)检测到病毒。在感染后第14天检测到由recPR8-H5N1毒株感染引起的特异性弱血清转化。结果,获得了一种具有改良特性的新型流感病毒株。