KARAKTERISASI DAN APLIKASI ANTIBODI MONOKLONALWDSSB5 UNTUK DETEKSI VIRUS DENGUE PADASEL C6/36 DENGAN METODE IMUNOSITOKIMIA

Abstract

Background: Dengue Fever (DF) and Dengue Hemorrhagic Fever (DHF) are caused by Dengue virus that consists of 4 serotypes Dengue 1, 2, 3 and 4. The clinical manifestations of Dengue infection in human have a broad spectrum ranging from asymptomatic to severe manifestations. Isolation of Dengue virus using cell culture C6/36 is considered as a gold standard for the diagnosis of Dengue virus infection. Dengue team of Gadjah Mada University successfully produced monoclonal antibodies of Dengue 3 virus originating from hybrid cell DSSC7, DSSE10 and WDSSB5. Detection of Dengue virus antigen in Ae. aegypti organ and in human blood smear with method imunocytochemical SBPC (Streptavidine Biotin Peroxide Complex) using DSSC7 primary antibody has high sensitivity and specificity, whereas WDSSB5 hybrid cell has not been characterized. The study aimed to identify characterization and application of WDSSB5 monoclonal antibody as primary antibody for detection of Dengue virus originating from serum of patiens positively carrying Dengue virus was inoculated in cell C6/36 using immunocytochemical SBPC method. Methods: The study was experimentally designed. Propagation of WDSSB5 hybridoma cell was performed in vitro and in vivo. The characterization consisted of classification test of WDSSB5 monoclonal antibody, examination of WDSSB5 acytes protein level, sensitivity and specificity of immunocytochemical SBPC method using WDSSB5 primary antibody and specificity of monoclonal antibody against Dengue antigen with Dot blot. Dengue virus originating from serum of patients positively carrying Dengue virus was inoculated in cell C6/36. Detection of Dengue virus antigen was made by using immunocytochemical SBPC method with WDSSB5 monoclonal antibody as primary antibody. Positive control was made using cell C6/36 infected with Dengue 1, 2, 3, 4 virus and inoculated in cell C6/36, whereas negative control uses cell C6/36 not infected with Dengue virus. Results: There was WDSSB5 monoclonal antibody. WDSSB5 monoclonal antibody belonged to class IgG and subclass IgG1. The least content of WDSSB5 monoclonal antibody that can detect Dengue antigen in cell C6/36 was 2.2 I¼g/I¼l. Sensitive WDSSB5 monoclonal antibody can detect Dengue 1, 2, 3, 4 virus antigen in cell C6/36 using immunocytochemical SBPC method. Conclusions: There was WDSSB5 monoclonal antibody specific of Dengue virus. WDSSB5 monoclonal antibody belonged to class IgG and subclass IgG1. WDSSB5 Monoclonal antibody can be applied to detection Dengue virus originating from serum of patients positively carrying Dengue virus was inoculated in cell C6/36 using imunositokimia SPBC method.