A群轮状病毒抗原检测试剂盒参考品的制备及稳定性考察  

Preparation and stability assessment of quality control reference materials for group A rotavirus antigen detection kits

作　　者：余娜 谢天宏[1] 戴永娟[1] 李华[1] 岳磊[1] 谢忠平[1] YU Na;XIE Tianhong;DAI Yongjuan;LI Hua;YUE Lei;XIE Zhongping(Institute of Medical Biology,Peking Union Medical College,Chinese Academy of Medical SciencesKuming Yunnan 650118,China)

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,云南昆明650118

出　　处：《中国病毒病杂志》2025年第1期88-93,共6页Chinese Journal of Viral Diseases

摘　　要：目的制备一套A群轮状病毒(group A rotavirus,RVA)抗原检测试剂盒(胶体金法)质量控制参考品,并对其稳定性进行考察。方法用细胞培养法制备的1组肠道病毒液、轮状病毒(rotavirus,RV)液及致腹泻的主要细菌培养物,建立RVA抗原检测试剂企业用参考品。阳性参考品(P1~P8)、阴性参考品(N1~N8)、检测限参考品S0、精密性参考品R0。随机抽取3套参考品(P1~P8、R0、S0),使用BioRad ChemiDoc MP系统将试剂盒条带亮度信号转化为灰度值,结果结合检测线(T线)灰度值的变异系数(coefficient of variation,CV)以及肉眼观察条带是否阳性且显色均一,考察分装均匀性;将参考品分别放置于4℃、25℃、37℃保存3、7 d及反复冻融2~6次,测定灰度值,利用Mann-Whitney U检验考察参考品的稳定性。结果阳性参考品、检测限参考品、精密性参考品显色条带均一,无肉眼可见的差异,灰度值CV在4%以内,参考品分装均匀性良好;冻融2~6次检测显色条带均一、无肉眼可见的差异,经灰度值检测及统计分析,差异无统计学意义(P>0.05);参考品在4℃、25℃及37℃放置7 d内,均具有较好的稳定性,差异无统计学意义(P>0.05)。结论本研究建立了RVA抗原检测试剂盒(胶体金法)企业用参考品,均一性、稳定性好,可用于RVA抗原检测试剂盒的质量控制。Objective To prepare a set of quality control reference materials for Group A rotavirus(RVA)antigen detection kits(colloidal gold method)and to evaluate their quality and stability.Methods A panel of intestinal virus preparations,rotavirus(RV)solutions,and cultures of major diarrhea-causing bacteria were established using cell culture methods to create reference materials for RVA antigen detection kits.Positive reference materials(P1~P8),negative reference materials(N1~N8),detection limit reference material(S0),and precision reference material(R0)were generated.Three sets of reference materials(P1~P8,R0,S0)were randomly selected,and the brightness signals of the kit strips were converted to grayscale values using the BioRad ChemiDoc MP system.The uniformity of the reference materials was assessed based on the coefficient of variation(CV)of grayscale values of the test line(T line)and visual inspection for uniform colouration.The reference materials were stored at 4℃,25℃,and 37℃for 3 and 7 days and subjected to 2–6 freeze-thaw cycles to evaluate their stability using grayscale value measurements and Mann-Whitney U tests.Results The positive reference materials,detection limit reference materials,and precision reference materials exhibited uniform color bands with no visible differences,and the CV of grayscale values was within 4%,indicating good uniformity.After 2-6 freezethaw cycles,the color bands remained uniform with no visible differences,and statistical analysis showed no significant differences(P>0.05).The reference materials demonstrated good stability when stored at 4℃,25℃,and 37℃for up to 7 days,with no significant differences observed(P>0.05).Conclusion Quality control reference materials for RVA antigen detection kits(colloidal gold method)are established with good uniformity and stability,which are suitable for quality control of RVA antigen detection kits.

关 键 词：A群轮状病毒 参考品 抗原检测试剂盒 胶体金 稳定性 

分 类 号：R373.25[医药卫生—病原生物学]