口蹄疫病毒抗体SA-ELISA快速检测方法的建立

doi:10.11843/j.issn.0366-6964.2023.05.023

Abstract

Abstract: In order to solve the problem of how to optimize and innovate the existing methods to establish a new method for FMD antibody detection, this study aimed to optimize the method based on liquid phase blocking ELISA to achieve rapid and sensitive detection of FMD virus antibody. In this study, a novel liquid phase blocking ELISA method (SA-LPBE) was established by combining the purification of IgG labeled biotin (dolphin anti-foot-and-mouth disease) from guinea pig serum with HRP labeled streptavidin (HRP-SA). SA-LPBE has a high consistency rate in bovine and sheep serum samples. Combined with the actual situation, the optimized serum antibody titer of cattle and sheep was determined to be ≥ 128, and that of pigs serum sample was determined to be ≥ 64. The results were determined to be positive, which was consistent with the results of commercial detection kits. And after compliance analysis, it has a compliance rate of 92.3% with the original kit, and the intra-batch variation coefficient is less than 5%, and the inter-batch variation coefficient is less than 10%. This method changed the original route of preparing rabbit anti-guinea pig antiserum and labeling HRP, improved the efficiency of enzyma-tic reaction, guaranteed the original sensitivity of the method and improved the specificity of the method; Using the advantages of quick reaction between biotin-labeled antibody and HRP-SA, stable labeling raw material and detection background value, the operation time of liquid phase blocking ELISA kit was shortened from 2 days to 3 hours, which solved the problems of tedious, long time, operation fatigue, instability and error-prone of the original operation. It can be applied to FMDV antibody detection, providing a technical method for FMD epidemic and clinical detection.

Key words: foot-and-mouth disease virus, liquid-phase blocking ELISA, enzyme-linked immunosorbent assay, antibody diagnosis, streptavidin

CLC Number:

S855.3

ZHOU Guangqing, LIU Xiaoqing, SHI Xijuan, YANG Dapeng, YUAN Ligang, CHANG Huiyun. Establishment of a Rapid Detection SA-ELISA Method for Anti Foot-and-Mouth Disease Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2020-2029.

References 30

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Establishment of a Rapid Detection SA-ELISA Method for Anti Foot-and-Mouth Disease Virus

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