title: Development of a lateral flow immunochromatographic assay for the rapid diagnosis of Orf virus infections.
abstract: A rapid and simple lateral-flow immunochromatographic assay (LFIA) was developed for the specific detection of Orf virus (ORFV) using two distinct monoclonal antibodies (MAbs: 5A5 and 6F2) against the ORFV ORF011 protein. The MAb 5A5 was conjugated with colloidal gold, and the MAb 6F2 and goat anti-mouse IgG were sprayed onto a nitrocellulose membrane in strips at positions designated test (T) and control (C), respectively. The results showed that samples of ORFV complexed with colloidal gold-conjugated MAb 5A5, were captured by MAb 6F2 at the T line resulting in the appearance of a purple band. When samples did not contain ORFV or when they contained a quantity of ORFV below the detection limit of the test, only the C line was visible. The analysis of sensitivity of the test demonstrated that the lowest detected quantity of ORFV was 2.03×10(3.0) TCID50/ml. Storage at room temperature for 6 months did not result in the loss of performance of the LFIA test. Using loop-mediated isothermal amplification (LAMP) as a reference test, the relative specificity and sensitivity of the LFIA test were determined to be 100% and 92.1%, respectively. Based on these results, the LFIA test developed may be a suitable tool for rapid on-site testing for ORFV infection.
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China.
authors.forename: Kui
authors.initials: K
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China.
authors.forename: Wenqi
authors.initials: W
authors.lastname: He
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China.
authors.forename: Jingying
authors.initials: J
authors.lastname: Bi
authors.affiliation: Laboratory Animal Center, Peking University, 5 Summer Palace Road, Beijing 100871, People's Republic of China.
authors.forename: Ximu
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China.
authors.forename: Di
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China.
authors.forename: Houshuang
authors.initials: H
authors.lastname: Huang
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China.
authors.forename: Yuexiang
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China.
authors.forename: Deguang
authors.initials: D
authors.lastname: Song
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China; Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis and Animal Research Center, Jilin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China. Electronic address: gaofeng@jlu.edu.cn.
authors.forename: Feng
authors.initials: F
authors.lastname: Gao
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods*; D003955:Diagnostic Tests, Routine / Q000379:methods*; D004474:Ecthyma, Contagious / Q000175:diagnosis*; D051379:Mice; D009923:Orf virus / Q000276:immunology*; D012680:Sensitivity and Specificity; D012756:Sheep
chemical_list: D000911:Antibodies, Monoclonal
keywords: Lateral flow immunochromatographic assay; Monoclonal antibodies; Orf virus; Rapid detection