Development and application of a colloidal gold test strip for detection of avian leukosis virus
- 297 Downloads
Avian leukosis virus (ALV) is an avian oncogenic retrovirus that induces leukemia-like proliferative diseases in chickens. ALV infection can result in the development of immunological tolerance and persistent viremia. Since effective vaccines against ALV are not yet available, its current prevention primarily depends on detection and eradication to establish exogenous ALV-free poultry flocks. In this study, a rapid and simple colloidal gold test strip method, specific for the group-specific antigen, p27 protein, was developed and systematically evaluated for the detection of ALV from different samples. The detection limit of this assay was as low as 6.25 ng/ml for p27 protein and 80 TCID50/ml for different subgroups of ALV. Besides, the test strip showed high specificity in the detection of different subgroups of ALV, including ALV-A, ALV-B, ALV-J, and ALV-K, with no cross-reaction with other avian pathogens. Furthermore, we artificially infected specific pathogen-free (SPF) chickens with ALV-J, collected cloacal swabs, and examined viral shedding using both test strips and ELISA. Results from the test strip were highly consistent with that from ELISA. In addition, 1104 virus isolates from anti-coagulant blood samples, 645 albumen samples, and 4312 meconium samples were tested, and the test strip results agreed with those of ELISA kit up to 97.1%. All the results indicated that the colloidal gold test strip could serve as a simple, rapid, sensitive, and specific diagnostic method for eradication of ALV in poultry farms.
KeywordsAvian leukosis virus Colloidal gold strip test Eradication ELISA Meconium
This study was supported by the National Key Research & Development (R&D) Plan (2016YFD0501605) and the National Natural Science Foundation of China (31761133002).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
All animal studies with chicken were approved by the Review Board of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences. All animal procedures were performed according to international standards on animal welfare.
- Adkins HB, Blacklow SC, Young JA (2001) Two functionally distinct forms of a retroviral receptor explain the nonreciprocal receptor interference among subgroups B, D, and E avian leukosis viruses. J Virol 758:3520–3526. doi: https://doi.org/10.1128/jvi.75.8.3520-3526.2001
- Du Y, Cui Z, Qin A (2000) Isolation of subgroup J avian leukosis viruses and their partial sequence comparison. Chin J Virol 2000(04):341–346. https://doi.org/10.13242/j.cnki.bingduxuebao.001274 (In Chinese)CrossRefGoogle Scholar
- Gao YL, Qin LT, Pan W, Wang YQ, Le Qi X, Gao HL, Wang XM (2010a) Avian leukosis virus subgroup J in layer chickens, China. Emerg Infect Dis 1610:1637–1638. https://doi.org/10.3201/eid1610.100780
- Gao YL, Shao HB, Luo QP, Pan W, Qin LT, Sun FF, Liu CN, Gao HL, Qi XL, Wang XM (2010b) Molecular epidemiology of avian leukosis virus isolates from some regions of China in 2009. Chin J Prev Vet Med 321:32–31. (In Chinese)Google Scholar
- Lai H, Zhang H, Ning Z, Chen R, Zhang W, Qing A, Xin C, Yu K, Cao W, Liao M (2011) Isolation and characterization of emerging subgroup J avian leukosis virus associated with hemangioma in egg-type chickens. Vet Microbiol 1513-4:275–283. https://doi.org/10.1016/j.vetmic.2011.03.037 CrossRefGoogle Scholar
- Payne LN, Brown SR, Bumstead N, Howes K, Frazier JA, Thouless ME (1991) A novel subgroup of exogenous avian leukosis virus in chickens. J Gen Virol 72 ( Pt 4)4:801–807. doi: https://doi.org/10.1099/0022-1317-72-4-801
- Payne LN, Howes K, Gillespie AM, Smith LM (1992) Host range of Rous sarcoma virus pseudotype RSV (HPRS-103) in 12 avian species: support for a new avian retrovirus envelope subgroup, designated J. J Gen Virol 73 (Pt 11)11:2995–2997Google Scholar
- Qiu Y, Qian K, Shen H, Jin W, Qin A (2011) Development and validation of an indirect enzyme-linked immunosorbent assay for the detection of avian leukosis virus antibodies based on a recombinant capsid protein. J Vet Diagn Investig 235:991–993. https://doi.org/10.1177/1040638711416966 CrossRefGoogle Scholar
- Sithigorngul W, Rukpratanporn S, Sittidilokratna N, Pecharaburanin N, Longyant S, Chaivisuthangkura P, Sithigorngul P (2007) A convenient immunochromatographic test strip for rapid diagnosis of yellow head virus infection in shrimp. J Virol Methods 1401-2:193–199. https://doi.org/10.1016/j.jviromet.2006.11.034 CrossRefGoogle Scholar
- Tsukamoto K, Hihara H, Kono Y (1991) Detection of avian leukosis virus antigens by the ELISA and its use for detection of infectious virus after cultivation of samples and partial characterization of specific pathogen-free chicken lines maintained in this laboratory. J Vet Med Sci 533:399–408CrossRefGoogle Scholar
- Venugopal K, Smith LM, Howes K, Payne LN (1998) Antigenic variants of J subgroup avian leukosis virus: sequence analysis reveals multiple changes in the env gene. J Gen Virol 79 ( Pt 4) sup1:757–766. doi: https://doi.org/10.1099/0022-1317-79-4-757
- Wang Q, Gao Y, Wang Y, Qin L, Qi X, Qu Y, Gao H, Wang X (2012) A 205-nucleotide deletion in the 3′ untranslated region of avian leukosis virus subgroup J, currently emergent in China, contributes to its pathogenicity. J Virol 8623:12849–12860. https://doi.org/10.1128/jvi.01113-12 CrossRefGoogle Scholar
- Zander DV, Raymond RG, McClary CF, Goodwin K (1975) Eradication of subgroups A and B lymphoid leukosis virus from commercial poultry breeding flocks. Avian Dis 193:403–423Google Scholar