title: Detection of dengue virus antigens in cultured cells by using protein A-gold-silver staining (pAgs) method.
abstract: A protein A-gold-silver (pAgs) staining was developed to detect dengue virus antigens in cultured cells. The method can be carried out in either newly-subcultured or monolayered cells. Dengue virus-inoculated C6/36 clone of Aedes albopictus cells and human endothelial cells appeared brown-yellowish color on the peripheral membrane of the infected cells. In many cases, the infected C6/36 cells appeared darker than that of the infected endothelial cells. The positive results from the inoculated C6/36 cells usually appeared as early as 2 days post-inoculation for types 1, 2, and 4 of dengue viruses and 3 days for the dengue 3 virus. The same batch of specimens detected by direct immunofluorescence antibody test (DFA) showed positive 4 days post-inoculation for the types 2, 3, and 4 of dengue viruses and 6 days for the dengue 1 virus. The result also showed that all pAgs-positive specimens were also DFA-positive, but not vice versa. It suggested that pAgs is not only sensitive but also specific for dengue virus detection from inoculated cultured cells.
authors.affiliation: Department of Parasitology, Kaohsiung Medical College, Taiwan.
authors.forename: W J
authors.initials: WJ
authors.lastname: Chen
authors.forename: S L
authors.initials: SL
authors.lastname: Chen
authors.forename: A H
authors.initials: AH
authors.lastname: Fang
authors.forename: M T
authors.initials: MT
authors.lastname: Wang
mesh_terms: D000330:Aedes / Q000276:immunology / Q000382:microbiology; D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D000956:Antigens, Viral / Q000276:immunology*; D001426:Bacterial Proteins; D002478:Cells, Cultured; D003716:Dengue Virus / Q000276:immunology*; D004730:Endothelium, Vascular / Q000382:microbiology; D005455:Fluorescent Antibody Technique; D017979:Gold Colloid; D006801:Humans; D007150:Immunohistochemistry; D009942:Organometallic Compounds; D016622:Silver Staining; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D001426:Bacterial Proteins; D017979:Gold Colloid; D009942:Organometallic Compounds; C070001:protein AG-gold complex