Development of an indirect ELISA against Orf virus using two recombinant antigens, partial B2L and F1L.

Autor: Zheng W; College of Animal Science, Guizhou University, Guiyang, China; Key Laboratory of Animal Diseases and Veterinary Public Health of Guizhou Province, College of Animal Science, Guizhou University, Guiyang, China., Zhang Y; College of Animal Science, Guizhou University, Guiyang, China; Key Laboratory of Animal Diseases and Veterinary Public Health of Guizhou Province, College of Animal Science, Guizhou University, Guiyang, China., Gu Q; College of Animal Science, Guizhou University, Guiyang, China; Key Laboratory of Animal Diseases and Veterinary Public Health of Guizhou Province, College of Animal Science, Guizhou University, Guiyang, China., Liang Q; College of Animal Science, Guizhou University, Guiyang, China; Key Laboratory of Animal Diseases and Veterinary Public Health of Guizhou Province, College of Animal Science, Guizhou University, Guiyang, China., Long Y; College of Animal Science, Guizhou University, Guiyang, China; Key Laboratory of Animal Diseases and Veterinary Public Health of Guizhou Province, College of Animal Science, Guizhou University, Guiyang, China., Wu Q; College of Animal Science, Guizhou University, Guiyang, China; Key Laboratory of Animal Diseases and Veterinary Public Health of Guizhou Province, College of Animal Science, Guizhou University, Guiyang, China., Xian S; College of Animal Science, Guizhou University, Guiyang, China; Key Laboratory of Animal Diseases and Veterinary Public Health of Guizhou Province, College of Animal Science, Guizhou University, Guiyang, China. Electronic address: xiansimei2005@163.com.
Jazyk: angličtina
Zdroj: Journal of virological methods [J Virol Methods] 2024 May; Vol. 326, pp. 114891. Date of Electronic Publication: 2024 Feb 07.
DOI: 10.1016/j.jviromet.2024.114891
Abstrakt: Orf is a highly contagious viral disease affecting goats and sheep. It is caused by Orf virus (ORFV) and has caused severe economic losses to the global goat industry, including in China. In this study, an indirect ELISA method for recombinant proteins based on truncated dominant antigenic epitopes of B2L and F1L genes of ORFV was established. A series of conditions and its performance were comprehensively evaluated. The optimized ELISA reaction conditions were: the optimal coating amount of antigen was 0.25 μg/mL, 5% skim milk powder was closed for 1 h, the optimal dilution of serum was 1:200, the optimal incubation time of the rabbit anti-goat IgG was 1:8000, the optimal color development time of TMB was 15 mins, and the threshold value of negative-positive was 0.358. The method specifically detects anti-ORFV antibodies and does not cross-react with positive sera for other common goat pathogenic bacteria antiserum. ORFV-positive sera were still positive after 1:512 dilution, with intra-batch coefficient of variation (CV) between 7.1% and 9.5% and inter-batch CV between 5.0% and 7.6%; 51% (92/180) of immunized goat serum samples were tested positive and 14.44% (14/63) of non-immunized goat serum samples were positive. The results show that the indirect ELISA antibody assay established in this study has good specificity, sensitivity and reproducibility, and provides a technical tool for clinical ORFV serum antibody detection and epidemiological investigation.
Competing Interests: Declaration of Competing Interest The authors(Weihao Zheng, You Zhang, Qinglin Gu, Qian Liang, Youci Long, Qin Wu, Simei Xian) declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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