Detection of hepatitis C virus specific core protein in serum of patients by a sensitive fluorescence enzyme immunoassay (FEIA)
| Autor: | Satoshi Tanaka, Atsumi Takata, Kendo Kiyosawa, Akira Hasegawa, Tomiko Kashiwakuma, Tadahiro Kajita, Michinori Kohara, Takeshi Tanaka, Nobu Hattori, Eiji Tanaka, Yohsuke Ohta, Hiroyuki Mori |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
medicine.drug_class
Hepatitis C virus Immunology Antibody Affinity Hepacivirus Antibodies Viral medicine.disease_cause Monoclonal antibody Immunofluorescence Virus Immunoenzyme Techniques Mice Antibody Specificity medicine BDNA test Animals Immunology and Allergy Hepatitis Antibodies Fluorescent Antibody Technique Indirect Mice Inbred BALB C medicine.diagnostic_test biology Viral Core Proteins Hepatitis Antigens Antibodies Monoclonal virus diseases Hepatitis C Virology Molecular biology digestive system diseases Titer Spectrometry Fluorescence Immunoassay biology.protein Female Antibody Epitope Mapping |
| Zdroj: | Journal of Immunological Methods. 190:79-89 |
| ISSN: | 0022-1759 |
| Popis: | A protein-capture fluorescence enzyme immunoassay (FEIA) was developed using monoclonal antibodies (mAbs) against recombinant hepatitis C virus (HCV) core protein. Four hybridoma cell lines (5E3, 5F11, 515S, 1080S) were established and characterized. These monoclonal antibodies (mAbs) each had IgG1 and IgG2 isotypes, and recognized major B cell epitopes within the immunodominant nucleoprotein amino terminal subregion. Using mAb 5F11 as the first antibody to the solid phase and β- d -galactosidase-conjugated mAb 5E3 as the second antibody to the protein, we established a specific HCV core protein capturing FEIA capable of detecting as little as 20 pg/ml of recombinant HCV core protein. HCV core protein in serum was detectable after treatment with 4.0% polyethyleneglycol, 0.5 M NaOH, and 5% Triton X-100. The results of a peptide inhibition assay indicated that this FEIA is specific for HCV RNA positive sera. The quantity of HCV core protein detected in serum was significantly correlated to the level of HCV RNA. The detection limit for HCV core proteins was an HCV RNA per titer of ∼ 10 4 /ml. Using this FEIA system, the detection ratio of HCV core protein in patients with chronic HCV infection was 92.3% ( 70 76 ). |
| Databáze: | OpenAIRE |
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