Detection of Dengue Virus Antigens in Cultured Cells by Using Protein A-Gold-Silver Staining (pAgs) Method

• Published in: MICROBIOLOGY and IMMUNOLOGY Vol. 37; no. 5; pp. 359 - 363
• Main Authors: Chen, Wei-June, Chen, Su-Lih, Fang, Ay-Huey, Wang, Ming-Tsan
• Format: Journal Article
• Language: English
• Published: Tokyo Blackwell Publishing Ltd 01.01.1993; Center For Academic Publications Japan; Center for Academic Publications Japan
• Subjects: Aedes - immunology; Aedes - microbiology; Aedes albopictus; Animals; Antibodies, Viral - analysis; Antigens, Viral - immunology; Bacterial Proteins; Biological and medical sciences; Cells, Cultured; Dengue virus; Dengue Virus - immunology; Endothelium, Vascular - microbiology; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; Gold Colloid; Humans; Immunohistochemistry; Microbiology; Organometallic Compounds; Protein A-gold-silver staining; Silver Staining; Staining and Labeling - methods; Techniques used in virology; Virology; Virus detection; Human; Cell culture; Endothelial cell; Immunogold labelling; Insecta; Protein A; Dengue virus; Flavivirus; Virus; Antigen; Aedes albopictus; Investigation method; Arthropoda; Culicidae; Infected cell; Flaviviridae; Invertebrata; Conjugated compound; Detection; Diptera
• ISSN: 0385-5600; 1348-0421
• DOI: 10.1111/j.1348-0421.1993.tb03222.x

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.