重配技术构建高产H1N1型流感疫苗病毒株

目的以经典重配技术制备高产H1N1流感疫苗病毒株。方法以野生型A1/云南昆明/03/2009(H1N1)作为HA及NA基因的供体株,以WHO疫苗株A/Perth/16/2009(H3N2)作为高产基因供体株,共同感染SPF鸡胚,经抗H3及抗N2血清中和筛选法及终末稀释法筛选高产重配H1N1病毒。结果获得一株重配H1N1流感病毒株,病毒血凝滴度为1∶4 096,病毒滴度为7.8 lg EID50/mL,显示为鸡胚高产病毒株;血凝抑制结果为1∶1 024,单向免疫扩散试验结果为阳性,证明抗原性与野生株一致;基因测序结果表明重配株的HA及NA基因序列与野生株序列一致。结论构建了高产重配H1H1流感疫苗病毒株,并应用经典重配技术建立了制备高产流感疫苗病毒株的技术平台。

Construction of high yield influenza H1N1 virus strain by reassortment

Objective To construct influenza H1N1 virus strain by reassortment. Methods SPF chick embryo were co-infected with influenza virus strain A1/Yunnan Kunming/03/2009 （ H1N1 ） and vaccine strain A/Perth/16/2009 （ H3N2 ） recommend by WHO. The reassortants were screened with antibody against strain A/Perth/16/2009 （H3N2） , subsequently subeultured in chick embryo. Results A chick embryo-adapted influenza HI N1 virus strain was obtained, of which the HI titer was maintained at 1 ： 1 024,the HA titer was 1 ： 4 096, and the virus titer was 7.8 lg EID50/mL, sequence analysis result showed that the reassortment virus has same HA and NA gene with the street virus. Conclusion A high yield influ- enza H1 N1 virus strain was constructed, and the plateform of producing influenza virus vaccine strain by the classical reas-sortment technique was established.