以重配技术制备H1N1流感病毒Vero细胞适应株  

Preparation of Vero Cell-Adapted Influenza H1N1 Virus Strain by Reassortment

作　　者：张英伟[1] 李卫东[1] 马娜[1] 姚忠萍[1] 燕景恩[1] 张勇[1] 刘雅灵[1] 杨佳[1] 吴忠香[1] 廖国阳[1] 杨净思[1] 李长贵[2] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所生物制品五室,昆明650118 [2]中国药品生物制品检定所疫苗三室,北京100050

出　　处：《中国生物制品学杂志》2009年第6期529-532,共4页Chinese Journal of Biologicals

基　　金：国家863科技计划项目(2006AA02Z409);云南省自然科学基金资助项目(2004C0720M;2006YX29);卫生部行业科研专项项目(200802023);云南省联合支持国家项目(2008GA007)

摘　　要：目的以重配技术制备H1N1流感病毒Vero细胞适应株,为以Vero细胞为基质生产流感疫苗奠定基础。方法以流感病毒Vero细胞适应株A/Yunnan/1/2005Va(H3N2)为母本株,与WHO推荐的2007～2008年度北半球流感疫苗生产用毒株A/Caledonia/20/99(H1N1)共同感染SPF鸡胚和Vero细胞,用抗A/Yunnan/1/2005Va(H3N2)抗体筛选重配病毒,在Vero细胞连续传12代,采用血凝抑制试验和RT-PCR鉴定病毒型别。将重配病毒经Vero细胞大量培养,重配前的病毒经鸡胚大量培养,分别经甲醛灭活,制备灭活疫苗,免疫ICR小鼠,检测其免疫原性。结果获得了1株Vero细胞适应的高产H1N1流感病毒株,连续传9代后,病毒血凝滴度维持在512,免疫原性与重配前的毒株相比,差异无统计学意义。结论通过流感病毒Vero细胞适应株与流行株的重配和抗体筛选,可以获得H1N1流感病毒Vero细胞适应株。Objective To prepare Vero cell-adapted influenza H1N1 virus strain by reassortment and lay a foundation of production of influenza vaccine using Vero cells as a substrate. Methods SPF chick embryo and Vero cells were co-infected with Vero cell-adapted influenza virus strain A / Yunnan / 1 / 2005Va （H3N2） and strain A / Caledonia / 20 / 99 （ H1N1 ） recommend by WHO for production of influenza vaccine in the Northern Hemisphere during 2007 - 2008. The reassortants were screened with antibody against strain A /Yunnan/l /2005Va （H3N2） subcuhured in Vero cells for 12 passages and identified for subtype by HI test and RT-PCR. The viruses before and after reassortment were cultured in a large quantity in chick embryos and Vero cells respectively, and inactivated with formaldehyde. ICR mice were immunized with the prepared inactivated influenza vaccines to observe the immunogenicities. Results A Vero cell-adapted influenza H1N1 virus strain was obtained, of which the HI titer was maintained at 512 after subculture for 9 passages, and the immunogenicity showed no significant difference with that before reassortment. Conclusion The Veto cell-adapted epidemic strain of influenza virus may be obtained by reassortment of Vero cell-adapted and epidemic strains.

关 键 词：流感病毒 H1N1亚型 遗传重配 VERO细胞 

分 类 号：R373.13[医药卫生—病原生物学]