中国兽医科学2025，Vol.55Issue(7)：891-897,7.DOI:10.16656/j.issn.1673-4696.2025.0134
牛轮状病毒间接免疫荧光试验方法的建立与初步应用
Establishment and preliminary application of indirect immunofluorescence detection method for bovine rotavirus
摘要
Abstract
Using the positive serum of bovine rotavirus(with a neutralization titer of 1∶256)as the primary antibody and the FITC-labeled goat anti-bovine IgG as the secondary antibody,an indirect immunofluorescence assay method for detecting bovine rotavirus was established.The working concen-trations and incubation times were optimized,and the optimal conditions were obtained.This method was used to detect bovine rotavirus in 48 clinical suspected samples,and the results were compared with those of two etiological detection methods,RT-PCR and virus isolation.The results showed that the op-timal working conditions were a 1∶300 dilution of the primary antibody and an incubation of 120 min-utes,a 1∶500 dilution of the secondary antibody and an incubation of 120 minutes;there was no cross-reaction with BCoV,IBRV,BVDV,BRSV,and BPIV3;when the virus titer was 101.25 TCID50/0.1 mL,there was still specific green fluorescence,and the fluorescence intensity was not affected by different times under the same conditions,the clinical sample detection results indicated that the IFA method had higher sensitivity than the virus isolation method,and the coincidence rate with the RT-PCR detec-tion results was 100％.The indirect immunofluorescence method established in this study has good specificity,sensitivity,repeatability,and accuracy,and can be used for the detection of clinical bovine rotavirus infection.关键词
牛/轮状病毒/间接免疫荧光法Key words
bovine/rotavirus/indirect immunofluorescence分类
农业科技引用本文复制引用
于文博,迟田英,郑辉,李博文,吴发兴,崔进,代飞燕..牛轮状病毒间接免疫荧光试验方法的建立与初步应用[J].中国兽医科学,2025,55(7):891-897,7.基金项目
云南省高原畜禽消化道营养与健康创新团队项目(202305AS350024) （202305AS350024）
云南省个旧市牛产业科技特派团项目(202304BI090007) （202304BI090007）
石林县农民院士科技服务站项目(A3012024017) （A3012024017）
