Multiple recombinant ELISA for the detection of bovine viral diarrhoea virus antibodies in cattle sera

Zoth, Silvina Chimeno; Taboga, Oscar

doi:10.1016/j.jviromet.2006.07.025

Cited by 37 publications

(28 citation statements)

References 28 publications

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“…The first generations of diagnostic ELISA kits utilized the extracts of virus infected cell cultures as ELISA detector antigens ( 14 , 15 ). Several ELISAs have been recently developed to detect BVDV infections using recombinant NS3 protein ( 16 - 19 ). Eukaryotic or prokaryotic recombinant NS3 is used for this purpose and this protein has shown high sensitivity and specificity to detect BVDV infection in comparison with whole virus antigen ( 16 , 20 ).…”

Section: Discussionmentioning

confidence: 99%

Simple Indirect Enzyme-Linked Immunosorbent Assay to Detect Antibodies Against Bovine Viral Diarrhea Virus, Based on Prokaryotically Expressed Recombinant MBP-NS3 Protein

Mahmoodi

Shapouri

Ghorbanpour

et al. 2014

Jundishapur J Microbiol

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Background:Bovine viral diarrhea (BVD) is an economically important disease of cattle distributed worldwide. Diagnosis of BVD relies on laboratory-based detection of its viral causing agent or virus specific antibodies and the most common laboratory method for this purpose is Enzyme-Linked Immunosorbent Assay (ELISA).Objectives:The current study was aimed to develop a simple indirect ELISA to detect antibodies against Bovine Viral Diarrhea Virus (BVDV) in the sera of infected cattle.Materials and Methods:A new simple indirect ELISA method was developed to detect BVDV infection by prokaryotically (Escherichia coli, BL21 strain) expressed recombinant whole nonstructural protein 3 (NS3) of BVDV (NADL strain). Four hundred bovine serum samples were evaluated by the newly developed NS3-ELISA and virus neutralization test (VNT) as the gold standard method to diagnose BVD. Among these samples, 289 sera had been previously tested by a commercial ELISA kit.Results:Statistical analyses showed a very high correlation between the results of the developed NS3-ELISA and VNT (kappa coefficient = 0.935, P < 0.001), with the relative sensitivity and specificity of 94% and 98.8%, respectively. There was also a high correlation between the results of NS3-ELISA and the commercial ELISA kit (kappa coefficient = 0.802, P < 0.001) with the relative sensitivity and specificity of 90.72% and 91.15%, respectively.Conclusions:The newly developed simple indirect ELISA showed high sensitivity and specificity with respect to VNT. Developing such a simple, sensitive, and specific ELISA which is much less expensive than the available commercial ELISA kits can improve the detection of BVDV infections, help to eliminate the disease from herds, and decrease economic losses caused by this disease.

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Section: Discussionmentioning

confidence: 99%

Simple Indirect Enzyme-Linked Immunosorbent Assay to Detect Antibodies Against Bovine Viral Diarrhea Virus, Based on Prokaryotically Expressed Recombinant MBP-NS3 Protein

Mahmoodi

Shapouri

Ghorbanpour

et al. 2014

Jundishapur J Microbiol

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“…In comparison with the complete virus, the recombinant proteins caused no infection and were easy to produce and purify. The replacement of the means of diagnosis of BVDV with E2 recombinant proteins used for the diagnosis of BVDV will have an extensive application prospects (Chimeno and Taboga, 2006). Note: Data with different superscripts stand for statistically significant difference (P < 0.01), and data with the same superscript stand for no statistically significant difference (P > 0.05).…”

Section: Discussionmentioning

confidence: 99%

Development of a sandwich Dot-ELISA for detecting bovine viral diarrhea virus antigen with E2 recombinant protein

Zhao

Zuo

Zhang

et al. 2009

Front. Agric. China

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The IgG antibodies of rabbit anti-E2 protein of the bovine viral diarrhea virus were prepared by a general method from high efficiency serum immunized by E2 recombinant protein antigen expressed in E. coli prokaryotic expression system and were labeled to make enzymelabeled antibody with the method of NaIO 4 . A sandwich Dot enzyme-linked immunosorbent assay (Dot-ELISA) for the detection of BVDV was developed. The optimal reaction conditions of Dot-ELISA were determined. The results show that optimal coating antibody was 300 μg$mL -1 , the working concentration of HRP-labeled antibody was 1∶50. The optimal blocking reagent and time were 5% bovine serum and 45 min. The minimum detection of the content of antigen reached 1.35 μg$mL -1 . Compared with the routine IDEXX ELISA test kit with the whole virus, its specificity, sensitivity and coincidence rate were 90.48%, 96.55% and 95.24%, respectively. Compared with the sandwich Dot-ELISA with the negative staining electron microscope and RT-PCR, the coincidence rates were 90.9% and 93.1%, respectively. In addition, Bovine viral diarrhea virus (BVDV) antigen of 178 samples collected from cow farms in the Hebei Province, China, were detected by the developed Dot-ELISA and the IDEXX BVDV antigen Test Kit simultaneously, BVDV antigen positive rate was 39.89%-41.01%. The result of detecting clinical samples demonstrated that the established method showed its specificity, sensitivity and repeatability, whereas the results were easily interpreted without an ELISA reader.

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“…Bovine Viral Diarrhea Virus is a member of the Pestivirus genus in the Flaviviridae family. Among viral proteins, Erns (previously E0 or gp48), E2 structural proteins and the non-structural NS3 protein have been identified as the principal immunogens of BVDV ( 8 ). Envelope glycoprotein Erns, encoding a dsRNase, that is capable of apoptosis and Interferon synthesis, is efficiently inhibited by the dsRNase ( 9 ).…”

Section: Introductionmentioning

confidence: 99%

Production of Monoclonal Antibody Against Recombinant Polypeptide From the Erns Coding Region of the Bovine Viral Diarrhea Virus

Shapouri

Ekhtelat

Abadi

et al. 2015

Jundishapur J Microbiol

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Background:Bovine viral diarrhea (BVD) is an economically important cattle disease with a worldwide distribution. Detection and elimination of animals persistently infected (PI) with bovine viral diarrhea virus (BVDV) is essential for the control of BVD and eradication of BVDV. There are usually no pathognomonic clinical signs of BVDV infection. Diagnostic investigations therefore rely on laboratory-based detection of the virus, or virus-induced antigens or antibodies.Objectives:Erns as an immunogenic protein of BVDV, is genetically and antigenically conserved among different isolates and therefore, is a candidate antigen for development of the enzyme linked immunosorbent assay (ELISA) for serological studies or identification of PI animals. The aim of this study was to produce a monoclonal antibody (MAb) against recombinant Erns.Materials and Methods:For this purpose, recombinant maltose-binding protein (MBP)-Erns protein was expressed in Escherichia coli and purified using amylose resin chromatography column and used as an antigen in MAb production. Spleen cells of the immunized mice with the recombinant antigen were fused with SP2/0 myeloma cells. Next, culture supernatants of primary clones of fused cells were screened by indirect ELISA. After three rounds of cloning, the reactivity of the MAbs with recombinant and natural antigen was established by Western blotting.Results:Based on our results, MAb against recombinant Erns was produced and reacted successfully with recombinant and natural antigens.Conclusions:With regards to the role of Erns in the identification of PI animals, it appears that Erns recombinant antigen and the specific monoclonal antibodies produced against it may be suitable for developing BVDV laboratory diagnostic assays.

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Multiple recombinant ELISA for the detection of bovine viral diarrhoea virus antibodies in cattle sera

Cited by 37 publications

References 28 publications

Simple Indirect Enzyme-Linked Immunosorbent Assay to Detect Antibodies Against Bovine Viral Diarrhea Virus, Based on Prokaryotically Expressed Recombinant MBP-NS3 Protein

Simple Indirect Enzyme-Linked Immunosorbent Assay to Detect Antibodies Against Bovine Viral Diarrhea Virus, Based on Prokaryotically Expressed Recombinant MBP-NS3 Protein

Development of a sandwich Dot-ELISA for detecting bovine viral diarrhea virus antigen with E2 recombinant protein

Production of Monoclonal Antibody Against Recombinant Polypeptide From the Erns Coding Region of the Bovine Viral Diarrhea Virus

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