Establishment of an indirect ELISA method based on VP6 protein for the detection of human group A rotavirus antibodies
10.3969/j.issn.1002-2694.2025.00.146
- VernacularTitle:人A组轮状病毒VP6蛋白间接ELISA抗体检测方法的建立
- Author:
Weibin ZHANG
1
;
Changkun LYU
;
Ying YANG
;
Mingyong WANG
;
Xiangpeng WANG
Author Information
1. 商丘医学高等专科学校医学技术学院,商丘 476100
- Publication Type:Journal Article
- Keywords:
human rotavirus group A;
VP6 protein;
indirect enzyme-linked immunosorbent assay;
antibody detection
- From:
Chinese Journal of Zoonoses
2025;41(9):932-938
- CountryChina
- Language:Chinese
-
Abstract:
This study established an indirect ELISA method for antibody detection based on human rotavirus group A(RVA)VP6 protein,to provide technical support for RVA antibody detection.An ELISA method for detecting RVA antibodies was established through a checkerboard assay for optimization of the reaction conditions with VP6 protein of RVA as the diagnostic antigen.Serum samples from healthy children and patients infected with RVA were collected and detected with the ELISA method.The optimal work-ing conditions for ELISA involved coating the ELISA plate with VP6 protein at a concentration of 2 μg/mL and diluting the serum at a ratio of 1∶250.The critical value of negative and positive samples was 0.137.The ELISA method had good specificity,sensitivity,and repeatability,and showed a 95%consistency rate with the western blotting antibody detection method.We tested 370 serum samples collected from children,which showed an antibody positivity rate of 81.1%.We additionally tested the acute and recovery phase sera from 15 patients infected with RVA,and observed a significantly higher RVA antibody titer in the recovery phase serum than the acute infection phase serum.The ELISA antibody detection method showed good specificity,sensitivity,and reproducibility,and therefore can be used for RVA antibody detection and auxiliary diagnosis of RVA infection.
