乙型肝炎病毒基因B型及基因C型两种国家标准品研制  

Preparation of National Standard for HBV genotype B and C

作　　者：李克坚[1] 郝晓甜 周诚[1] LI Kejian;HAO Xiaotian;ZHOU Cheng(National Institute for the Control of Pharmaceutical and Biological Products,Beijing,China,100050)

机构地区：[1]中国食品药品检定研究院传染病诊断一室,北京100050

出　　处：《分子诊断与治疗杂志》2024年第1期113-117,共5页Journal of Molecular Diagnostics and Therapy

基　　金：国家科技重大专项项目(2018ZX10732401)。

摘　　要：目的研制乙型肝炎病毒(HBV)基因B型及基因C型两种国家标准品。方法收集我国不同地区的HBV感染者血浆样品,经HBV五项、DNA病毒检测,从中筛选出11份基因分型标准品候选品样品。经HBV序列测定,进化树分析,初步确定2份样品分别为HBV B型及C型标准品候选品。分别将2份候选品离心、分装后,进行HBV基因分型验证测定,并进行定量的协作标定测定。结果确认筛选出HBV基因B型及基因C型标准品,其HBV DNA浓度分别为4.67×10~7IU/mL(CV为3.6%)及3.66×10^(8)IU/mL(CV为2.9%)。装量检查,大于0.5 mL/支,符合规定。均匀性检查,两者P值分别为0.428(F值为1.140)及0.420(F值为1.173),分装的标准品支与支之间浓度均匀无差异。将两种标准品在不同方式保存(反复冻融、4℃保存、室温及37℃放置)后,测定结果与-80℃保存样品测定结果相比较,绝对偏差均在±0.2的范围内,方差分析P值均大于0.1;另外,-20℃保存12个月后与-80℃保存样品两者测定数据分析,B型P值为0.237(F值为1.934)、C型P值为0.173(F值为2.737),P值均大于0.1,稳定性验证符合规定。结论制备了HBV基因B型及基因C型两种国家标准品,为我国HBV基因分型试剂的质量控制和标准化提供了依据。Objective To develop two kinds of national standard hepatitis B virus B genotype and C genotype.Metheods A total of 11 genotyping standard candidate samples were screened from plasma samples of HBV infected patients in different regions of China.Two samples were identified as HBV type B and C standard samples by HBV sequence determination and evolutionary tree analysis.The two candidates were centrifuged and packaged,respectively,for HBV genotyping validation and quantitative collaborative calibration.Results Confirm and screen HBV B genotype and C genotype standard samples.The virus content of B genotype and C genotype standard samples was 4.67×107 IU/mL(CV 3.6%)and 3.66×108 IU/mL(CV 2.9%).he loading volume is more than 0.5 mL/piece,which is in line with the regulations.The P values of the two were 0.428(F=1.140)and 0.420(F=1.173),respectively.There was no difference in the concentration between branches.After the two standards were stored in different ways(repeated freeze-thawing,4℃storage,room temperature and 37℃storage),the results were compared with those of the samples stored at-80℃.The absolute deviations were within the range of±0.2,and the P values of ANOVA were all greater than 0.1.In addition,after 12 months stored at-20℃and stored at-80℃,the P-value of type B was 0.237(F value was 1.934),and the P-value of type C was 0.173(F value was 2.737),both P values were greater than 0.1.The stability verification was in line with the requirements.Conclusion Two kinds of national standard HBV genotype B and C genotypes were prepared,which provided the basis for the quality control and standardization of HBV genotyping reagent in China.

关 键 词：乙型肝炎病毒 基因型 国家标准品 定量测定 

分 类 号：R512.62[医药卫生—内科学]