Enzyme-linked immunosorbent assay (ELISA) is often
used to test bovine leukemia virus (BLV) infection. However, commercially
available kits test in South America detect only antibodies against the gp51
protein. With the aim to improve the sensitivity of the test, we developed here
a two-step indirect dual ELISA test that included both proteins p24 and gp51,
expressed and produced in E. coli and
baculovirus expression system respectively. Two hundred ten BLV sera, stated as
double positive or double negative by the combination of commercial agar gel
immunodiffusion (AGID) assay and a gp51-ELISA test, were tested with our in
house dual rp24/rgp51 ELISA. Firstly, we checked the purified, optimized and
standardized proteins as antigen by the checkerboard technique, and set up our
in house ELISA test. The concordance
correlation coefficient (CCC) and coefficient of variation (CV) intraplate
repeatability levels were within the values established by the international
standards. The statistical analysis demonstrated the value of sera correctly
ranked highest (93.48%), and for 0.3 cutoff, the sensitivity was 95.65% and the
specificity 91.30%. In conclusion, the rp24/rgp51 ELISA developed and
standardized here demonstrated to have good analytical characteristics to be
considered for screening of BLV.
References
[1]
Lucas, M.H. (1992) Viral Diseases. Enzootic Bovine Leukosis. In: Andrews, A.H., Blowey, R.W., Boyd, H. and Eddy, R.G., Eds., Bovine Medicine: Diseases and Husbandry, 2nd Edition. Wiley-Blackwell, Hoboken, 530-537.
[2]
Robert, D.H., Lucas, M.H. and Wibberley, G. (1986) To Compare the Incubation Period Following Intratracheal and Subcutaneous Inoculation of Bovine Leucosis Virus Infected Lymphocytes and to Study Their Clearance from the Circulation. Veterinary Immunology and Immunopathology, 11, 351-358.
https://doi.org/10.1016/0165-2427(86)90037-1
[3]
Gonzalez, E.T., Oliva, G.A., Valera, A.R., Bonzo, E., Licursi, M. and Etcheverrigaray, M.E. (2001) Leucosis Enzoótica Bovina: Evaluación De Técnicas Diagnósticas (ID, ELISA-1, WB, PCR) En Bovinos Inoculados Experimentalmente. Analecta Veterinaria, 21, 12-20.
[4]
Brenner, J., Van-Haam, M., Savir, D. and Trainin, Z. (1989) The Implication of BLV Infection in the Productivity, Reproductive Capacity and Survival Rate of a Dairy Cow. Veterinary Immunology and Immunopathology, 22, 299-305.
https://doi.org/10.1016/0165-2427(89)90017-2
[5]
Monti, G.E., Frankena, K. and De Jong, M.C.M. (2007) Evaluation of Natural Transmission of Bovine Leukaemia Virus within Dairy Herds of Argentina. Epidemiology and Infection, 135, 228-237. https://doi.org/10.1017/S0950268806006637
[6]
Weiss, R.A. (2006) The Discovery of Endogenous Retroviruses. Retrovirology, 3, 67.
https://doi.org/10.1186/1742-4690-3-67
[7]
Troiano, L.D.C., Thomaz-Soccol, V., Agottani, J.V.B., Brodzinski, J., Penha, T.R., and Ozaki, S.C. (2013) Production, Characterization, and Use of Monoclonal Antibodies Against gp51 Protein to Diagnose Bovine Leukemia Virus Infection. BioResearch Open Access, 2, 55-60. https://doi.org/10.1089/biores.2012.0295
[8]
González, E.T., Licursi, M., Oliva, G.A. and Etcheverrigaray, M.E. (1999) Diagnóstico De La Leucosis Enzoótica Bovina (LEB) En Animales Naturalmente Infectados. XIV Congresso Estadual De Medicina Veterinaria, III Congreso De Medicina Veterinaria Do Cone Sul. Gramado, Brasil. Proceeding, 192.
[9]
Monti, G.E., Frankena, K., Engel, B., Buist, W., Tarabla, H.D. and de Jong, M.C. (2005) Evaluation of a New Antibody-Based Enzyme-Linked Immunosorbent Assay for the Detection of Bovine Leukemia Virus Infection in Dairy Cattle. Journal of Veterinary Diagnostic Investigation, 17, 451-457.
https://doi.org/10.1177/104063870501700507
[10]
Kittelberger, R., Reichel, M.P., Meynell, R.M, Tham, K.M. and Molloy, J.B. (1999) Detection of Antibodies against the Core Protein p24 of the Bovine Leukaemia Virus in Cattle for Confirmatory Serological Testing. Journal of Virological Methods, 77, 109-114. https://doi.org/10.1016/S0166-0934(98)00143-8
[11]
Heeney, J.L., Valli, V.E. and Montesanti J. (1988) Alterations in Humoral Immune Response to Bovine Leukaemia Virus Antigens in Cattle with Lymphoma. Journal of General Virology, 69, 659-666. https://doi.org/10.1099/0022-1317-69-3-659
[12]
Simard, C., Richardson, S., Dixon, P., Bélanger, C. and Maxwell, P. (2000) Enzyme-Linked Immunosorbent Assay for the Diagnosis of Bovine Leukosis: Comparison with the Agar Gel Immunodiffusion Test Approved by the Canadian Food Inspection Agency. Canadian Journal of Veterinary Research, 64, 101-106.
[13]
Mammerickx, M., Potetelle, D., Bruck, C. and Burny, A. (1984) Diagnosis of Enzootic Bovine Leukosis with an Immunossay Test (Elisa) Utilizing a Monoclonal Antibody Ann. MedVet Medical, 123, 55-63.
[14]
Crowther, J.R. (2009) Tritation of Reagents. In: Crowther, J.R., Ed., The ELISA Guidebook (Methods in Molecular Biology), 2nd Edition. Humana Press, New York City, 79-109.
[15]
Sanchez, J., Dohoo, I.R., Markham, F., Leslie, K. and Conboy, G. (2002) Evaluation of the Repeatability of a Crude Adult Indirect Ostertagia Ostertagi ELISA and Methods of Expressing Test Results. Veterinary Parasitology, 109, 75-90.
https://doi.org/10.1016/S0304-4017(02)00194-2
[16]
OIE. (2008) Enzootic Bovine Leukosis. In: World Organisation for Animal Health, Ed., Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, World Organisation for Animal Health, Paris, Volume 2, Chapter 2.4.11, 729-738.
[17]
Xu, H., Lohr, J. and Greiner, M. (1997) The Selection of ELISA Cut-Off Points for Testing Antibody to Newcastle Disease by Two-Graph Receiver Operating Characteristic TG-ROC Analysis. Journal of Immunological Methods, 208, 61-64.
https://doi.org/10.1016/S0022-1759(97)00128-2
[18]
Sagata, N., Yasunaga, T., Tsuzuku-Kawamura, J., Ohishi, K., Ogawa, Y. and Ikawa, Y. (1985) Complete Nucleotide Sequence of the Genome of Bovine Leukemia Virus: Its Evolutionary Relationship to Other Retroviruses. Proceedings of the National Academy of Sciences of the United States of America, 82, 677-681.
https://doi.org/10.1073/pnas.82.3.677
[19]
González, E.T., Oliva, G.A., Ando, Y. and Etcheverrigaray, M.E. (1988) Bovine Leukemia Virus. Antigen Production and Serologic Detection. 2nd International Conference on the Impact of Viral Diseases on the Development of Latin American Countries and the Caribbean Region, Mar del Plata, Argentina.
[20]
Van der Maaten, M. and Miller, J.M. (1990) Bovine Leukosis Virus. In: Dinter, Z. and Morein, B., Eds., Virus Infections of Ruminants, Elsevier Science Publishers, Amsterdam, 419-429. https://doi.org/10.1016/b978-0-444-87312-5.50058-8
[21]
De Giuseppe, A., Feliziani, F., Rutili, D. and De Mia, G.M. (2004) Expression of the Bovine Leukemia Virus Envelope Glycoprotein (gp51) by Recombinant Baculovirus and Its Use in an Enzyme-Linked Immunosorbent Assay. Clinical and Diagnostic Laboratory Immunology, 11, 147-151.
https://doi.org/10.1128/cdli.11.1.147-151.2004
[22]
OIE. (2013) Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. In: OIE, Ed., Principles and Methods of Validation of Diagnostic Assays for Infectious Diseases, Chapter 1.1.6. (NB: Version adopted in May 2013).
[23]
Pelzer, K.D. (1997) Economics of Bovine Leukemia Virus Infection. The Veterinary Clinics of North America. Food Animal Practice, 13, 129-141.
https://doi.org/10.1016/S0749-0720(15)30368-6
