| A群轮状病毒抗原检测试剂盒的研究及性能评价 |
| 投稿时间：2024-09-05 修订日期：2024-10-23 点此下载全文 |
| 引用本文：余娜,谢天宏,李华,戴永娟,岳磊,吴晋元,谢忠平.A群轮状病毒抗原检测试剂盒的研究及性能评价[J].医学研究杂志,2025,54(3):142-146, 186 |
| DOI：
10.11969/j.issn.1673-548X.2025.03.026 |
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| 中文摘要: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1×104pfu/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1×107pfu/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。 |
| 中文关键词:轮状病毒 免疫胶体金技术 性能评价 快速检测 |
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| Research and Performance Evaluation of Group A Rotavirus Antigen Detection Kit. |
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| Abstract: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 1×104pfu/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 1×107pfu/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, epidemiological research, and vaccine strain screening. It has good application prospects and practical value. |
| keywords:Rotavirus Immunocolloidal gold technique Performance evaluation Rapid detection |
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