福建农业学报2025，Vol.40Issue(7)：649-655,7.DOI:10.19303/j.issn.1008-0384.2025.07.001
鸭短喙矮小综合征病毒抗体间接ELISA检测方法的建立与初步应用
An Indirect ELISA for Diagnosing Duck Short Beak and Dwarfism Syndrome
摘要
Abstract
[Objective]A rapid serological method was developed for detecting antibodies against the Short Beak and Dwarf Syndrome Virus(SBDSV)in ducks.[Methods]The epidemic strain M15 of SBDSV was purified by sucrose density gradient centrifugation and inactivated to be used as the solid-phase coating antigen.After optimization of reaction conditions,a highly specific indirect enzyme-linked immunosorbent assay(ELISA)was developed and verified by testing for assay specificity,repeatability,and accuracy on clinic specimens for detecting the specific antibody in ducks.[Results]The optimal ELISA analytical conditions included the uses of antigen at the concentration of 0.1 mg·mL-1,coating the antigen at 37℃for 2 h followed by incubation overnight at 4℃and blocked with 1%BSA at 37℃for 2 h,serum dilution at the rate of 1∶200,and 0.3214 as the positive/negative cutoff point.The newly developed assay exhibited high specificity with no cross-reactivity on other common waterfowl viruses.It was highly sensitive as shown by remaining positive on hyperimmune serum(with an LPAI titer of 28)up to 1∶204 800 dilution.The repeatability on the intra-and inter-assays was high with coefficients of variation below 6%.On 70 serum specimens of ducks suspected of SBDSV infection,the assay delivered 90%of the results agreed with those obtained by LPAI.On the 648 serum samples of 1-day-old ducklings submitted for testing in Fujian,an antibody positivity rate of approximately 48.30%was detected by the new assay indicating some degrees of SBDSV infection on the birds at duck farms in the province.[Conclusion]The indirect ELISA for detecting SBDSV serum antibodies newly developed in this study is highly specific and suitable for large-scale serological testing,which could facilitate a rapid and effective monitoring means on epidemic SBDSV in the country and,in general,provide a technic support for the detection of the viral infection in ducklings.关键词
鸭短喙矮小综合征/间接ELISA/抗体检测Key words
short beak and dwarfism syndrome/indirect ELISA/antibody detection分类
农业科技引用本文复制引用
朱小丽,王劭,董慧,程晓霞,江丹丹,肖世峰,陈少莺,陈仕龙..鸭短喙矮小综合征病毒抗体间接ELISA检测方法的建立与初步应用[J].福建农业学报,2025,40(7):649-655,7.基金项目
福建省科技计划公益类专项(2022R1026006、2022R1026004、2021R1026008、2023R1024003) （2022R1026006、2022R1026004、2021R1026008、2023R1024003）
福建省农业高质量发展超越"5511"协同创新工程项目(XTCXGC2021018) （XTCXGC2021018）
福建省农业科学院对外合作项目(DWHZ2024-05) （DWHZ2024-05）
中央引导地方科技发展专项(2022L3019) （2022L3019）
福建省农业科学院科技创新团队建设项目(CXTD2021034) （CXTD2021034）
国家自然科学基金项目(32302868) （32302868）
