Early Detection of goats infected with Lentivirus Small Ruminant virus by ELISA assay

800x600 Normal 0 21 false false false ES-AR X-NONE X-NONE MicrosoftInternetExplorer4 Many established protocols are available to produce antigens for ELISA tests but expensive equipments, the purification methods or recombinant technologies are not always available to veterinary diagnostic laboratories. The aim of this work was to develop an alternative ELISA test to detect antibodies against Lentivirus Small Ruminant Virus (Caprine arthritis-encephalitis virus-CAEV) in goats, simple to elaborate with a low cost of production. The antigen was obtain from a whole cellular lysates of goat synovial membrane cells (GSMC) infected with CAEV and treated with SDS 0,1%. The whole-viral antigen was capable of detecting antibodies against several viral proteins including the p28 kDa (viral capsid), the p33 kDa, the p44 kDa (capsid precursors) and p97 kDa (transmembrane glycoprotein). The comparison with others assays as indirect immunofluorescence showed a high correlation with the ELISA results and the polimerasse chain reaction (PCR) demonstrated that most of the goat seropositives by ELISA, had proviral DNA in the milk cells. In conclusion, the viral antigen obtained from GSMC was simple to elaborate, low cost of production. The ELISA test was capable of detecting antibodies against CAEV, to detect early infection.

[1]  C. van Maanen,et al.  Diagnostic performance of ELISA and PCR in identifying SRLV-infected sheep and goats using serum, plasma and milk samples and in early detection of infection in dairy flocks through bulk milk testing. , 2010, Veterinary microbiology.

[2]  Silvia Inês Sardi,et al.  Produção de antígeno viral para o sorodiagnóstico da artrite-encefalite caprina utilizando um teste imunoenzimático (ELISA) , 2009 .

[3]  C. van Maanen,et al.  Evaluation of Five Enzyme-Linked Immunosorbent Assays and an Agar Gel Immunodiffusion Test for Detection of Antibodies to Small Ruminant Lentiviruses , 2007, Clinical and Vaccine Immunology.

[4]  Silvia Inês Sardi,et al.  Isolamento e identificação do vírus da Artrite encefalite caprina, a partir do co-cultivo de células mononucleares do sangue com células de membrana sinovial de cabra , 2006 .

[5]  D. Klein,et al.  Diagnostic tests for small ruminant lentiviruses. , 2005, Veterinary microbiology.

[6]  D. Klein,et al.  Transmission of small ruminant lentiviruses. , 2004, Veterinary microbiology.

[7]  R. Leite,et al.  A Labelled Avidin–Biotin ELISA to Detect Antibodies to Caprine Arthritis-encephalitis Virus in Goats' Sera , 1999, Veterinary Research Communications.

[8]  R. Schoborg.  Analysis of caprine arthritis encephalitis virus (CAEV) temporal gene expression in infected cells. , 2002, Virus research.

[9]  S. Rosati,et al.  Genetic Heterogeneity of Small Ruminant Lentiviruses Involves Immunodominant Epitope of Capsid Antigen and Affects Sensitivity of Single-Strain-Based Immunoassay , 2002, Clinical and Vaccine Immunology.

[10]  M. Lara,et al.  IDENTIFICAÇÃO IMUNO-SOROLÓGICA DE ANTICORPOS ANTI-VÍRUS DA ARTRITE-ENCEFALITE DOS CAPRINOS: COMPARAÇÃO DAS TÉCNICAS DE IMUNODIFUSÃO EM GEL DE ÁGAR, ENSAIO IMUNOENZIMÁTICO E IMUNOFLUORESCÊNCIA INDIRETA , 2002 .

[11]  V. Moojen,et al.  Imunofluorescência utilizando isolados brasileiros no diagnóstico sorológico de infecção por lentivírus em caprinos , 2002 .

[12]  V. Moojen,et al.  Detecção da infecção pelo vírus da artrite-encefalite caprina: imunodifusão em ágar e reação em cadeia da polimerase com "primers" degenerados , 2001 .

[13]  Roberto Soares de Castro,et al.  Lentivírus de pequenos ruminantes (CAEV e Maedi-Visna): revisão e perspectivas , 2001 .

[14]  P. Singh,et al.  Simple and Rapid Method for Production of Whole-Virus Antigen for Serodiagnosis of Caprine Arthritis-Encephalitis Virus by Enzyme-Linked Immunosorbent Assay , 2001, Clinical Diagnostic Laboratory Immunology.

[15]  E. Peterhans,et al.  B-cell epitopes of the envelope glycoprotein of caprine arthritis-encephalitis virus and antibody response in infected goats. , 2000, The Journal of general virology.

[16]  I. Hötzel,et al.  Immune response to caprine arthritis-encephalitis virus surface protein induced by coimmunization with recombinant vaccinia viruses expressing the caprine arthritis-encephalitis virus envelope gene and caprine interleukin-12. , 2000, Vaccine.

[17]  Y. Chebloune,et al.  Ovine lentivirus-infected macrophages mediate productive infection in cell types that are not susceptible to infection with cell-free virus. , 1999, The Journal of general virology.

[18]  S. Valas,et al.  Caprine arthritis-encephalitis virus in semen of naturally infected bucks , 1999 .

[19]  J. Costa,et al.  MÉTODOS LABORATORIAIS PARA O DIAGNÓSTICO DA INFECÇÃO PELO VÍRUS DA IMUNODEFICIÊNCIA HUMANA (HIV) , 1998 .

[20]  Roberto Soares de Castro,et al.  Produção de antígeno nucleoprotéico do vírus da artrite-encefalite caprina e comparação com o do vírus Maedi-Visna para utilização em teste de imunodifusão em ágar gel , 1998 .

[21]  E. Peterhans,et al.  Serologic diagnosis of ovine lentiviruses by whole virus ELISA and AGID test. , 1998, Zentralblatt fur Veterinarmedizin. Reihe B. Journal of veterinary medicine. Series B.

[22]  K. Olsson,et al.  A Long Term Study of Goats Naturally Infected with Caprine Arthritis-Encephalitis Virus , 1996, Acta Veterinaria Scandinavica.

[23]  J. Kwang,et al.  Serological diagnosis of caprine lentivirus infection by recombinant immunoassays , 1995 .

[24]  J. Thorsen,et al.  Chemiluminescent detection of caprine arthritis encephalitis virus with a PCR-generated single stranded nonradiolabelled probe. , 1995, Veterinary microbiology.

[25]  T. Besser,et al.  Evaluation of a Kinetic Enzyme-Linked Immunosorbent Assay for Detection of Caprine Arthritis-Encephalitis Virus-Specific Antibodies , 1994, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[26]  W. Heneine,et al.  Detection of caprine arthritis-encephalitis virus by polymerase chain reaction , 1993, Journal of clinical microbiology.

[27]  J. Clements,et al.  Nucleotide sequence and transcriptional analysis of molecular clones of CAEV which generate infectious virus. , 1990, Virology.

[28]  E. Peterhans,et al.  Detection of antibodies to caprine arthritis-encephalitis virus by protein G enzyme-linked immunosorbent assay and immunoblotting , 1989, Journal of clinical microbiology.

[29]  D. Knowles,et al.  Precursor polypeptides of caprine arthritis-encephalitis lentivirus structural proteins. , 1988, The Journal of general virology.

[30]  D. S. Adams,et al.  Antigenic cross-reactivity between caprine arthritis-encephalitis, visna and progressive pneumonia viruses involves all virion-associated proteins and glycoproteins. , 1985, The Journal of general virology.

[31]  J. Clements,et al.  Activation of caprine arthritis-encephalitis virus expression during maturation of monocytes to macrophages , 1983, Infection and immunity.