ABSTRACT
Epstein-Barr virus (EBV) is the main oncogenic lymphotropic agent of the Herpesviridae family and is globally distributed. EBV acute infection occurs in young adults producing infectious mononucleosis. Detection of anti-viral capside antigen (VCA) antibodies indicates previous or present EBV infection. Moreover, high titles of anti-VCA antibodies are observed in EBV-associated neoplasic disorders, such as lymphomas in AIDS patients. The objective of this study was the development and optimization of P3HR1 cell slides for the EBV serologic detection by indirect immunofluorescence (IIF) assay. P3HR1 exponential growth culture cells were stimulated with phorbol-12-mirystoil-13-acetate, collected at different time points and used for slide preparation. IIF assay was performed in each slide using an anti-EBV positive serum as primary antibody. An 11% increase in VCA expression was observed at 40 hours post-stimulation. Data was confirmed by Western blot and immunodetection. Intra- and inter-lot precisions of the developed slides were evaluated for IgG and IgM antibodies using EBV-positive sera and positive samples for other members of the Herpesviridae family. Neither false-positive or false negative results were obtained for EBV detection nor cross-reaction was observed with other members of the Herpesviridae family with the developed slides. In conclusion, the slides here presented can be a useful instrument for acute EBV infection diagnosis and for the serologic detection of IgG anti-VCA antibodies in EBV-associated neoplastic disorders.
Subject(s)
Cell Culture Techniques/instrumentation , Cell Line, Tumor/immunology , Epstein-Barr Virus Infections/diagnosis , Herpesvirus 4, Human/isolation & purification , Adult , Antigens, Viral/analysis , Antigens, Viral/immunology , Burkitt Lymphoma/immunology , Capsid Proteins/analysis , Capsid Proteins/immunology , Epstein-Barr Virus Infections/immunology , Equipment Design , Fluorescent Antibody Technique, Indirect , Herpesvirus 4, Human/immunology , Humans , Sensitivity and SpecificityABSTRACT
Epstein-Barr virus (EBV) is the main oncogenic lymphotropic agent of the Herpesviridae family and is globally distributed. EBV acute infection occurs in young adults producing infectious mononucleosis. Detection of anti-viral capside antigen (VCA) antibodies indicates previous or present EBV infection. Moreover, high titles of anti-VCA antibodies are observed in EBV-associated neoplasic disorders, such as lymphomas in AIDS patients. The objective of this study was the development and optimization of P3HR1 cell slides for the EBV serologic detection by indirect immunofluorescence (IIF) assay. P3HR1 exponential growth culture cells were stimulated with phorbol-12-mirystoil-13-acetate, collected at different time points and used for slide preparation. IIF assay was performed in each slide using an anti-EBV positive serum as primary antibody. An 11
increase in VCA expression was observed at 40 hours post-stimulation. Data was confirmed by Western blot and immunodetection. Intra- and inter-lot precisions of the developed slides were evaluated for IgG and IgM antibodies using EBV-positive sera and positive samples for other members of the Herpesviridae family. Neither false-positive or false negative results were obtained for EBV detection nor cross-reaction was observed with other members of the Herpesviridae family with the developed slides. In conclusion, the slides here presented can be a useful instrument for acute EBV infection diagnosis and for the serologic detection of IgG anti-VCA antibodies in EBV-associated neoplastic disorders.