Expression of the Bovine Leukemia Virus Envelope Glycoprotein (gp51) by Recombinant Baculovirus and Its Use in an Enzyme-Linked Immunosorbent Assay

ABSTRACT The gene encoding the major envelope glycoprotein (gp51) with its signal sequence, represented by an additional NH2-terminal 33-residue amino acid sequence of bovine leukemia virus (BLV), was inserted into a baculovirus transfer vector. A recombinant virus expressing a secreted gp51 protein in insect cells was isolated. The recombinant gp51 expressed was characterized by using an anti-BLV monoclonal antibody by both Western blotting analysis and enzyme-linked immunosorbent assay (ELISA). The secreted gp51 was used as an antigen, and an ELISA with recombinant gp51 (rgp51) was developed for the detection of BLV antibodies. This new procedure was compared with a previous ELISA method for the detection of BLV antibodies and an agar gel immunodiffusion test performed with an unpurified BLV antigen preparation. The comparative testing of field samples showed that the ELISA with rgp51 is more specific and also suitable for the testing of pooled sera.

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Agar gel immunodiffusion analysis using baculovirus-expressed recombinant bovine leukemia virus envelope glycoprotein (gp51/gp30T-)

Journal of Veterinary Science ◽

10.4142/jvs.2009.10.4.331 ◽

2009 ◽

Vol 10 (4) ◽

pp. 331 ◽

Cited By ~ 6

Author(s):

Seong In Lim ◽

Wooseog Jeong ◽

Dong Seob Tark ◽

Dong Kun Yang ◽

Chang Hee Kweon

Keyword(s):

Envelope Glycoprotein ◽

Bovine Leukemia Virus ◽

Leukemia Virus ◽

Virus Envelope ◽

Agar Gel ◽

Immunodiffusion Analysis

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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 ◽

10.1177/104063870501700507 ◽

2005 ◽

Vol 17 (5) ◽

pp. 451-457 ◽

Cited By ~ 16

Author(s):

Gustavo E. Monti ◽

Klaas Frankena ◽

Bas Engel ◽

Willem Buist ◽

Héctor D. Tarabla ◽

...

Keyword(s):

Maximum Likelihood ◽

Dairy Cattle ◽

Immunosorbent Assay ◽

Bovine Leukemia Virus ◽

Conditional Independence ◽

Latent Class ◽

Leukemia Virus ◽

Latent Class Model ◽

Enzyme Linked Immunosorbent Assay ◽

Conditional Dependence

The objective of this study was to validate a new blocking enzyme-linked immunosorbent assay (ELISA) (designated M108 for milk and S108 for serum samples) for detecting bovine leukemia virus (BLV) infection in dairy cattle. Milk, serum, and ethylenediaminetetraacetic acid–blood samples were collected from 524 adult Holstein cows originating from 6 dairy herds in Central Argentina. The M108 and S108 were compared with agar gel immunodiffusion (AGID), polymerase chain reaction and a commercial ELISA. Because there is currently no reference test capable of serving as a gold standard, the test sensitivity (SE) and specificity (SP) were evaluated by the use of a latent class model. Statistical inference was performed by classical maximum likelihood and by Bayesian techniques. The maximum-likelihood analysis was performed assuming conditional independence of tests, whereas the Bayesian approach allowed for conditional dependence. No clear conclusion could be drawn about conditional dependence of tests. Results with maximum likelihood (under conditional independence) and posterior Bayes (under conditional dependence) were practically the same. Conservative estimates of SE and SP (with 95% confidence intervals) for M108 were 98.6 (96.7; 99.6) and 96.7 (92.9; 98.8) and for S108 99.5 (98.2; 99.9) and 95.4 (90.9; 98.1), respectively. The ELISA 108 using either milk or serum to detect BLV-infected animals had comparable SE and SP with the official AGID and a commercial ELISA test, which are currently the most widely accepted tests for the serological diagnosis of BLV infection. Therefore, ELISA 108 can be used as an alternative test in monitoring and control programs.

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