Immunohistochemical detection of porcine reproductive and respiratory syndrome virus using colloidal gold.
Two cytopathic agents were isolated on porcine alveolar macrophages following inoculation with homogenates of lung tissues from pigs showing respiratory problems. These isolates were identified as porcine reproductive and respiratory syndrome (PRRS) virus isolates by indirect immunofluorescence using a PRRS virus (PRRSV) specific monoclonal antibody (MAb) and were designated as LHVA-92-1 and LHVA-92-2. Immunogold electron microscopy using a porcine PRRS positive serum pool and protein A-gold resulted in an intense labelling of aggregates of viral particles. Dark specific cytoplasmic staining of porcine alveolar macrophages infected with both virus isolates could be observed by immunogold silver staining (IGSS) using the specific MAb. This method proved effective in detecting PRRSV antigens in several ethanol-fixed tissues of piglets intranasally inoculated with the supernatants of macrophages infected with each isolate. Immunogold silver staining was also successfully used for the detection of PRRSV antigens on sections of formalin-fixed paraffin-embedded lung tissues and on frozen sections of lungs. The present results indicate that colloidal gold may be useful for the identification and immunohistochemical detection of PRRSV in tissues.
Magar
R
R
Laboratoire d'Hygiène Vétérinaire et Alimentaire, Agriculture Canada, Saint-Hyacinthe, Québec.
Larochelle
R
R
Robinson
Y
Y
Dubuc
C
C
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D018018:Arterivirus / Q000276:immunology / Q000302:isolation & purification* / Q000648:ultrastructure; D018174:Arterivirus Infections / Q000382:microbiology / Q000662:veterinary*; D002478:Cells, Cultured; D003588:Cytopathogenic Effect, Viral; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D008168:Lung / Q000382:microbiology* / Q000473:pathology; D016676:Macrophages, Alveolar / Q000382:microbiology; D016253:Microscopy, Immunoelectron; D012141:Respiratory Tract Infections / Q000382:microbiology / Q000662:veterinary*; D013047:Specific Pathogen-Free Organisms; D013552:Swine; D013553:Swine Diseases / Q000382:microbiology*; D013577:Syndrome
D000956:Antigens, Viral