HIV-1标准株资源库的初步建立与应用  

Primary construction and application of National bank for HIV-1 standard strains

作　　者：任莉[1] 何林[1] 朱美玲 王彦[1] 孔德生[1] 王琛[1] 冯毅[1] 邵一鸣 邢辉[1] 马春涛[1] 马丽英[1] Ren Li;He Lin;Zhu Meiling;Wang Yan;Kong Desheng;Wang Chen;Feng Yi;Shao Yiming;Xing Hui;Ma Chuntao;Ma Liying(National Center for AIDS/STD Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]中国疾病预防控制中心性病艾滋病预防控制中心,北京102206

出　　处：《中华实验和临床病毒学杂志》2023年第3期280-285,共6页Chinese Journal of Experimental and Clinical Virology

基　　金：国家"十三五"重大传染病防治科技重大专项(2018ZX10734404-005)。

摘　　要：目的构建代表我国HIV-1主要流行亚型的标准株,建立种子库和工作库。方法按照《病原微生物菌(毒)种标准株艾滋病病毒毒株建立技术规范》要求,复核鉴定分析病毒的基因型和表型。对6株HIV-1标准株进行逐级放大扩增培养,构建第二代种子库和第三代的工作库,液氮冻存。结果获得了6株HIV-1标准株,其中3株为CRF07BC(NRPC2.4.9003、NRPC2.4.9005、NRPC2.4.9006),1株为CRF01AE(NRPC2.4.9001),1株为CRF08BC(NRPC2.4.9002),1株为URF(NRPC2.4.9004)。表型检测6株病毒均为CCR5嗜性、非融合诱导型。TCID_(50)均大于1×10^(5)/ml,p24抗原浓度均高于2 ng/ml,构建了每株不少于20支的种子库以及每株不少于50支的工作库,应用HIV-1标准株对抗病毒药物PEG2kC34、PEG5kC34、LP-19以及中和抗体LSEVh-LS-F进行评价。结论获得了涵盖我国主要流行亚型的HIV-1标准株6株,建立了HIV-1标准株资源库。填补了我国HIV-1标准株保藏的空白,为药物和疫苗评价提供了技术支撑。Objective To construct standard strains representing the main epidemic clades of HIV-1 in China,amplify the virus strains,and establish a seed lot.Methods Six isolates of HIV-1 virus were identified and analyzed in genotype and phenotype,according to"interpretation for the social organization of the Standard strains of pathogenic microorganism-technical specifications for establishment of HIV strains".The isolates were amplified and cultivated to generate the secondary generation primary seed lot and the third generation working seed lot as frozen storage in liquid nitrogen.Results Six HIV-1 standard strains were obtained,of which 3 strains are CRF_07BC(NRPC2.4.9003,NRPC2.4.9005,NRPC2.4.9006),1 strain is CRF_01AE(NRPC2.4.9001),1 strain is CRF_08BC(NRPC2.4.9002),and 1 strain is URF(NRPC2.4.9004).Phenotypic detection showed that all six strains are CCR5 tropics and Non syncytia inducing virus.TCID_(50)were all greater than 1×10^(5)/ml,and concentrations of p24 antigen were all higher than 2 ng/ml.A primary seed lot with no less than 20 vials per strain and a working seed lot with no less than 50 vials per strain were constructed.The standard virus strains were used in evaluating antiviral drugs PEG2kC34,PEG5kC34,LP-19,and neutralizing antibody LSEVh LS-F.Conclusions Six standard strains of HIV-1 virus covering the three main epidemic subtypes of HIV-1 in China have been obtained,and a storage of HIV-1 standard strain was constructed.It meets the need of the preservation of HIV-1 standard strains in China and provides support for drug and vaccine evaluation.

关 键 词：人免疫缺陷病毒 国家标准株 病毒扩增 资源库 应用 

分 类 号：R373[医药卫生—病原生物学]