Detection of bovine leukemia virus specific antibodies using recombinant p24-ELISA.

Animals; Antibodies, Viral/blood; Cattle; Cloning, Molecular; Enzootic Bovine Leukosis/blood/diagnosis/immunology; Enzyme-Linked Immunosorbent Assay/veterinary; Escherichia coli/metabolism; Gene Expression Regulation; Leukemia Virus, Bovine/immunology; Recombinant Proteins; Viral Core Proteins/immunology

Abstract :

[en] We developed an indirect ELISA test (rp24-ELISA) for the detection of antibodies against the p24 capsid protein of bovine leukemia virus (BLV) in bovine serum samples. A recombinant p24 (rp24) was expressed in the Escherichia coli expression system, purified in a nickel charged resin and used as antigen in the ELISA test. A cut-off point was established considering the distribution of reactivity values obtained by rp24-ELISA on a set of 555 field serum samples that were stated as double positive (DP) or double negative (DN) by the combination of commercial agar gel immunodiffusion (AGID) assay and gp51-ELISA tests results. The rp24-ELISA showed good concordance with the agar gel immunodiffusion assay, when 710 serum samples were analyzed. In addition, rp24-ELISA demonstrated to be a precise assay with good repeatability and reproducibility and a better analytical sensitivity than AGID. From 67 discordant rp24-ELISA-AGID sera, 4 negative reactors were from the DP group, 28 positive samples were from the DN group and 35 positive samples were stated as negative by AGID but positive by the gp51-ELISA. Samples from this last subgroup were sent to an OIE reference laboratory where 28 samples were stated as negative, 5 samples were stated as positive and 2 as inconclusive. Complete concordance with blind previous results from an international proficiency test panel confirmed the capability of the assay to discriminate between infected and non-infected animals. In conclusion, the rp24-ELISA developed and standardized demonstrated to have good analytical characteristics to be considered for screening of BLV.

Disciplines :

Veterinary medicine & animal health

Author, co-author :

Gutierrez, G.

Alvarez, I.

Fondevila, N.

Politzki, R.

Lomonaco, M.

Rodriguez, Sabrina ; National Institute of Agriculture Technology INTA-Castelar, Argentina > CNIA > Institute of Virology

Dus Santos, M. J.

Trono, K.

Language :

English

Title :

Detection of bovine leukemia virus specific antibodies using recombinant p24-ELISA.

Publication date :

12 June 2009

Journal title :

Veterinary Microbiology

ISSN :

0378-1135

eISSN :

1873-2542

Publisher :

Elsevier, Netherlands

Volume :

137

Issue :

3-4

Pages :

224-34

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 23 January 2013

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