基于8E5细胞的HIV-1病毒载量能力验证质控品制备及评估
Preparation and evaluation of proficiency testing materials for HIV-1 viral load capacity based on 8E5 cells
梅朋飞 1米云婷 1朱禹静 2周德 3邓煜川 4金聪 1姚均 1邢文革 1潘品良 1吕毅1
作者信息
- 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
- 2. 宿迁市第一人民医院
- 3. 云南省妇幼保健院
- 4. 北京市疾病预防控制中心
- 折叠
摘要
目的 制备及评价基于8E5细胞的人类免疫缺陷病毒(human immunodeficiency virus 1,HIV-1)病毒载量能力验证质控品的均一性、稳定性和反复冻融性.方法 将8E5细胞上清原液10倍梯度稀释得到浓度依次为106、105、104、103拷贝/ml的4组HIV-1病毒载量能力验证质控品,应用病毒载量分析仪进行检测,将病毒拷贝数其换算成lg值,计算均值((x))、标准差(s)、变异系数(coefficient of variation,CV)、变化趋势,并对质控品的均一性、稳定性和反复冻融性进行评价.结果 在4个浓度梯度的均一性评价中,各浓度组质控品的CV分别为1.2％、2.6％、1.8％和3.2％.稳定性评价实验中,在-20 ℃条件下,各组质控品在5个月评价期间内,检测结果不随时间延长而改变(P＞0.05).在4 ℃条件下,各组质控品在5周评价期间内,检测结果随时间延长而未改变(P＞0.05).在25℃条件下,各组质控品在15d评价期间内,检测结果随时间延长而减少(P＜0.05).冻融次数影响评价中,反复冻融次数对质控品影响不大,其检测结果均在((x)±2s)范围浮动.结论 基于8E5细胞上清液制备的质控品,在-20 ℃和4 ℃保存条件下均一性和稳定性较好,可作为HIV-1病毒载量能力验证质控品.
Abstract
Objective This study aims to prepare and evaluate the homogeneity,stability,and repeatability of HIV-1 viral load proficiency testing materials(PTMs)based on 8E5 cells.Methods Four groups of proficiency testing materials for HIV-1 viral load with concentrations of 106,105,104,and 103copies/ml were obtained by 10-fold gradient dilution of 8E5 cell supernatant,and the viral load analyzer was applied to detect the viral copy numbers and its conversion into lg value.The mean((x)),standard deviation(s),coefficient of variation(CV),and trend were calculated to evaluate the homogeneity,stability,and repeatability of PTMs.Results In the homogeneity evaluation experiment of the four concentration gradients,the CVs of the PTMs in each concentration group were 1.2％,2.6％,1.8％,and 3.2％,respectively.In the stability evaluation experiment,the detection results did not change with the prolongation of time during the 5-month evaluation period for each group of PTMs at-20 ℃(P＞0.05).Meanwhile,the results did not change with time during the 5-week evaluation period for each group of PTMs at 4 ℃(P＞0.05).However,the results decreased with time during the 15-day evaluation period for each group of PTMs at 25 ℃(P＜0.05).In evaluating the effect of freeze-thaw times,the number of repeated freeze-thaws had little effect on the PTMs,whose detection results fluctuated within the range of((x)±2s).Conclusions In this study,the PTMs based on 8E5 cells had good homogeneity and stability under the storage conditions of-20 ℃ and 4 ℃ and could be used as a quality control for HIV-1 viral load capacity validation.
关键词
人类免疫缺陷病毒/病毒载量/RNA/8E5细胞/能力验证Key words
Human immunodeficiency virus 1/Viral load/RNA/8E5 cells/Proficiency testing引用本文复制引用
基金项目
北京市自然科学基金(7204282)
国家科技重大专项(20152X10001001-002)
出版年
2024
NETL
NSTL
万方数据