title: Development of an Indirect ELISA for Serological Diagnosis of Bovine herpesvirus 5.
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.
authors: Laboratório de Bacteriologia, Centro de Desenvolvimento Tecnológico - Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors: Luana A
authors: LA
authors: Dummer
authors: Laboratório de Bacteriologia, Centro de Desenvolvimento Tecnológico - Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors: Itauá L
authors: IL
authors: Araujo
authors: Laboratório de Virologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
authors: Fabrício S
authors: FS
authors: Campos
authors: Laboratório de Bacteriologia, Centro de Desenvolvimento Tecnológico - Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors: Matheus C
authors: MC
authors: da Rosa
authors: Laboratório de Imunologia Aplicada, Centro de Desenvolvimento Tecnológico - Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors: Paula F
authors: PF
authors: Finger
authors: Laboratório de Bacteriologia, Centro de Desenvolvimento Tecnológico - Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors: Patricia D
authors: PD
authors: de Oliveira
authors: Laboratório de Imunologia Aplicada, Centro de Desenvolvimento Tecnológico - Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors: Fabricio R
authors: FR
authors: Conceição
authors: Laboratório de Virologia e Imunologia Animal, Faculdade de Veterinária, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors: Geferson
authors: G
authors: Fischer
authors: Laboratório de Virologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.|Fundação Estadual de Pesquisa Agropecuária, Saúde Animal - Instituto de Pesquisas Veterinárias Desidério Finamor (IPVDF), Eldorado do Sul, RS, Brazil.
authors: Paulo M
authors: PM
authors: Roehe
authors: Laboratório de Bacteriologia, Centro de Desenvolvimento Tecnológico - Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors: Fábio P L
authors: FP
authors: Leite
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D019540:Area Under Curve; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000821:virology; D004285:Dogs; D018792:Encephalitis, Viral / Q000175:diagnosis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005843:Geography; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000276:immunology; D025881:Herpesvirus 5, Bovine; D061985:Madin Darby Canine Kidney Cells; D008590:Meningoencephalitis / Q000175:diagnosis* / Q000276:immunology; D009500:Neutralization Tests; D010843:Pichia; D012372:ROC Curve; D011994:Recombinant Proteins / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014759:Viral Envelope Proteins / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C091563:glycoprotein D, bovine herpesvirus 5