A群轮状病毒抗原检测试剂盒的研究及性能评价  

Research and Performance Evaluation of Group A Rotavirus Antigen Detection Kit

作　　者：余娜 谢天宏[1] 李华[1] 戴永娟[1] 岳磊[1] 吴晋元[1] 谢忠平[1] YU Na;XIE Tianhong;LI Hua(Institute of Medical Biology,Peking Union Medical College,Chinese Academy of Medical Sciences,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,昆明650118

出　　处：《医学研究杂志》2025年第3期142-146,186,共6页Journal of Medical Research

摘　　要：目的通过制备A群轮状病毒(group A rotavirus,RVA)抗原检测试剂盒并对其性能进行评价,为轮状病毒(rotavirus,RV)的快速检测提供有效手段。方法采用胶体金纳米粒子(gold nanoparticles,AuNPs)标记Rabbit anti-G1P[8]Ab1,将Goat anti-G1P[8]Ab2及Goat Anti-Rabbit IgG分别包被在硝酸纤维膜(nitrocellulose membrane,NC膜)的检测线(T线)和质控线(C线)上,将吸水纸、NC膜、结合垫、样品垫依次组装到PVC底板上,切割成3.75毫米/条,安装到适配卡壳,即制成RVA抗原检测试剂盒;采用轮状病毒抗原检测国家参考品(简称“国家参考品”)和本室制备企业参考品,分别对试剂盒敏感度、特异性、包容性、高剂量钩状效应、稳定性、重复性等性能进行考察,同时用同类产品对比评价。结果敏感度:用国家参考品L考察,最低检出限为1∶128倍稀释,相当于企业参考品最低检出限浓度1×10^(4)pfu/ml或2344copies/ml的病毒含量水平,且与同类商品化产品比较,敏感度高8倍左右;特异性:本试剂盒与10种临床常见腹泻致病病原体均不发生交叉反应;包容性:本试剂盒能同时检测G1~G4、G8、G9等多种不同血清型RVA;重复性:本试剂盒重复性检测考察,条带颜色均一,一致性较好;高剂量钩状效应:在1×10^(7)pfu/ml范围内不存在高剂量钩状效应;热加速稳定性:分别在37℃、50℃条件下保存14天,试剂盒检测条带清晰且均匀。结论本研究制备了性能良好的RVA抗原检测试剂盒,适用于临床辅助诊断、流行病学研究及疫苗毒株筛选等多方面快速检测工作,具有良好的应用前景及实用价值。Objective To prepare a group A rotavirus(RVA)antigen detection kit and evaluates its performance,providing an effective means for rapid detection of rotavirus(RV).Methods In this study,colloidal gold nanoparticles(AuNPs)were used to label Rabbit anti-G1P[8]Ab1.The Goat anti-G1P[8]Ab2 and Goat Anti-Rabbit IgG were respectively coated on the detection line(T line)and quality control line(C line)of nitrocellulose membrane(NC membrane).The absorbent paper,NC membrane,binding pad,and sample pad were assembled onto a PVC substrate in sequence and cut into 3.75mm strips.They were then installed into an adapter cartridge to establish a RVA antigen detection kit.National reference standards for rotavirus antigen detection and reference standards prepared by our laboratory were used to investigate the sensitivity,specificity,inclusiveness,high-dose hook effect,stability,repeatability,and compare and evaluate similar testing kits available for sale.Results Sensitivity:when tested with national reference substance L,the lowest detection limit is diluted 1∶128 times,which was equivalent to the virus content level of the lowest detection limit concentration of enterprise reference substance 1×10^(4)pfu/ml or 2344copies/ml,the sensitivity was about 8 times higher than that of similar commercial products.Specificity:there was no cross-reaction between this kit and 10 common clinical pathogens causing diarrhea.Inclusiveness:this kit could simultaneously detect multiple different serotypes of RVA,including G1-G4,G8,G9,etc.Repeatability:this reagent kit was tested repeatedly,and the color of the bands was uniform with good consistency.High-dose hook effect:there was no high-dose hook effect within the range of 1×10^(7)pfu/ml.Thermal acceleration stability:after being stored at 37℃and 50℃for 14days,the test strip of the kit was clear and uniform.Conclusion This study has developed a high-performance RVA antigen detection kit,which is suitable for rapid detection in various aspects such as clinical auxiliary diagnosis,epidemi

关 键 词：轮状病毒 免疫胶体金技术 性能评价 快速检测 

分 类 号：R331[医药卫生—人体生理学]