家畜生态学报2026，Vol.47Issue(4)：85-93,9.DOI:10.3969/j.issn.1673-1182.2026.04.012
ASFV p30蛋白间接ELISA抗体检测方法的建立与分析软件开发应用
Development of an Indirect ELISA for ASFV p30 Protein Antibody Detection and Its Software Application
摘要
Abstract
This study aimed to establish an indirect enzyme-linked immunosorbent assay(ELISA)based on the African swine fever virus(ASFV)p30 protein and to achieve automated analysis of test re-sults.Eukaryotically expressed no-Tag p30 protein was used as the solid-phase antigen.After optimizing ELISA reaction conditions using a checkerboard titration method,the assay's specificity and sensitivity were validated.Software for automated ELISA data analysis was coded in Python.The established indirect ELISA was then applied to detect ASFV antibodies in 700 serum samples collected from various swine populations in a city of Jiangsu Province in 2017 and from 2020 to 2024.The optimal conditions were deter-mined as follows:coating with 0.125 μg/mL antigen at 4℃ for 16 h,blocking with 10 g/L gelatin at 37℃for 2 h,incubation with serum diluted 1∶100 at 37℃ for 20 min,incubation with secondary antibody di-luted 1∶10 000 at 37℃ for 20 min,and color development in the dark at 27℃ for 8 min,with a cut-off value of 0.4.Compared to a commercial kit,the total coincidence rate of the developed method was 97.4％,with a sensitivity of 100％and a specificity of 96.4％.Results generated by the ELISA software were con-sistent with manual calculations.The overall positive rate for ASFV antibodies in the clinical serum sam-ples was 6.14％.All samples from 2017 and 2024 were negative,whereas positive samples were detected each year from 2020 to 2023,with annual positive rates ranging from a minimum of 1.61％to a maximum of 16.70％.The established method demonstrates high specificity and sensitivity,and the accompanying software provides reliable automated result analysis.This integrated approach offers a practical tool for clinical ASFV antibody detection,supporting rapid diagnosis and effective disease control.关键词
非洲猪瘟/p30蛋白/间接ELISA/抗体检测/软件开发应用Key words
African Swine Fever/p30 protein/indirect ELISA/antibody detection/software develop-ment and application分类
农业科技引用本文复制引用
朱恒志,马清霞,肖琦,张冯禧,李娜,赖虹桥,李子轩,宋京泽,解建平,温立斌,何孔旺..ASFV p30蛋白间接ELISA抗体检测方法的建立与分析软件开发应用[J].家畜生态学报,2026,47(4):85-93,9.基金项目
泰州市"凤城英才计划"113生物医药专项(评审类人才项目)(泰人才办[2022]23号) （评审类人才项目）
2025亚夫科技服务项目KF(25)1213 （25）
山东省生猪产业技术体系疫病控制岗位(SDAIT-08-07) （SDAIT-08-07）
