Research and Performance Evaluation of Group A Rotavirus Antigen Detection Kit
10.11969/j.issn.1673-548X.2025.03.026
- VernacularTitle:A群轮状病毒抗原检测试剂盒的研究及性能评价
- Author:
Na YU
1
;
Tianhong XIE
1
;
Hua LI
1
Author Information
1. 650118 昆明,中国医学科学院/北京协和医学院医学生物学研究所
- Publication Type:Journal Article
- Keywords:
Rotavirus;
Immunocolloidal gold technique;
Performance evaluation;
Rapid detection
- From:
Journal of Medical Research
2025;54(3):142-146,186
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare a group A rotavirus(RVA)antigen detection kit and evaluates its performance,providing an effec-tive means for rapid detection of rotavirus(RV).Methods In this study,colloidal gold nanoparticles(AuNPs)were used to label Rab-bit 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 car-tridge to establish a RVA antigen detection kit.National reference standards for rotavirus antigen detection and reference standards pre-pared 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 suit-able 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.
