Development of an Indirect ELISA for Serological Diagnosis of Bovine herpesvirus 5

Luana A Dummer; Itauá L Araujo; Fabrício S Campos; Matheus C da Rosa; Paula F Finger; Patricia D de Oliveira; Fabricio R Conceição; Geferson Fischer; Paulo M Roehe; Fábio P L Leite

doi:10.1371/journal.pone.0149134

Development of an Indirect ELISA for Serological Diagnosis of Bovine herpesvirus 5

PLoS One. 2016 Feb 11;11(2):e0149134. doi: 10.1371/journal.pone.0149134. eCollection 2016.

Affiliations

¹ Laboratório de Bacteriologia, Centro de Desenvolvimento Tecnológico - Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
² Laboratório de Virologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
³ Laboratório de Imunologia Aplicada, Centro de Desenvolvimento Tecnológico - Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
⁴ Laboratório de Virologia e Imunologia Animal, Faculdade de Veterinária, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
⁵ Fundação Estadual de Pesquisa Agropecuária, Saúde Animal - Instituto de Pesquisas Veterinárias Desidério Finamor (IPVDF), Eldorado do Sul, RS, Brazil.

Abstract

Bovine herpesviruses 1 and 5 (BoHV-1 and BoHV-5) are economically important pathogens, associated with a variety of clinical syndromes, including respiratory and genital disease, reproductive failure and meningoencephalitis. The standard serological assay to diagnose BoHV-1 and BoHV-5 infections is the virus neutralization test (VNT), a time consuming procedure that requires manipulation of infectious virus. In the present study a highly sensitive and specific single dilution indirect ELISA was developed using recombinant glycoprotein D from BoHV-5 as antigen (rgD5ELISA). Bovine serum samples (n = 450) were screened by VNT against BoHV-5a and by rgD5ELISA. Compared with the VNT, the rgD5ELISA demonstrated accuracy of 99.8%, with 100% sensitivity, 96.7% specificity and coefficient of agreement between the tests of 0.954. The rgD5ELISA described here shows excellent agreement with the VNT and is shown to be a simple, convenient, specific and highly sensitive virus-free assay for detection of serum antibodies to BoHV-5.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Viral / blood
Area Under Curve
Cattle
Cattle Diseases / diagnosis*
Cattle Diseases / virology
Dogs
Encephalitis, Viral / diagnosis*
Encephalitis, Viral / immunology
Enzyme-Linked Immunosorbent Assay / methods*
Geography
Herpesviridae Infections / diagnosis*
Herpesviridae Infections / immunology
Herpesvirus 5, Bovine
Madin Darby Canine Kidney Cells
Meningoencephalitis / diagnosis*
Meningoencephalitis / immunology
Neutralization Tests
Pichia
ROC Curve
Recombinant Proteins / chemistry
Reproducibility of Results
Sensitivity and Specificity
Viral Envelope Proteins / immunology
Viral Envelope Proteins / isolation & purification*

Substances

Antibodies, Viral
Recombinant Proteins
Viral Envelope Proteins
glycoprotein D, bovine herpesvirus 5

Grants and funding

This research was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (grants 478461/2004-6 and 140287/2010-8). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.