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      An ELISA method using serum derived HDAg for the sorological detection of HDV antigens and antibodies Translated title: Método imunoenzimático usando antígeno delta derivado do soro para detecção sérica de antígeno e anticorpo do vírus da hepatite delta

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          Abstract

          One of the main difficulties related to the detection of the Hepatitis Delta Virus (HDV) antigen and antibody has been the source of the needed HD antigen since HDV containing human and animal livers are very difficult to obtain and since yield is low. This fact prompted us to try to use the serum of patients in the acute phase of HDV infection as a source of HDAg and turn to enzyme immunoassays (EIA) instead of RIA for the sake of easiness and economy in the amount of HDAg needed. The antigen for EIA was obtained from patients during the acute phase of HDV infection and the antibody from patients who have been carriers for many years. For the detection of the antigen, a sandwich type method was employed, whereas for the antibody a competition assay was developed. In order to assess the relative specificity and sensibility of the test, the antibody assay was compared to a commercial RIA (C. RIA, Abbott) and to a non-commercial RIA (NC RIA). Forty-two sera were tested by the two methods and only in two cases discrepant results were obtained. Its is concluded that: 1) sera from patients in the acute and chronic phases of HDV infection can be used as source of both antigen and antibody, for immunoassays; 2) EIA and RIA have comparable relative specificity and sensibility and 3) EIA is easier to perform, cheaper, non-hazardous, has a longer shelf-life and saves scarce HDAg.

          Translated abstract

          Um dos maiores problemas no desenvolvimento de metodologia para detecção de antígenos e anticorpos do vírus da hepatite delta (VHD) tem sido a fonte de antígenos, uma vez que fígados humanos e de animais infectados pelo VHD são de difícil obtenção e baixo rendimento Ao uso de soro de pacientes na fase aguda da infecção pelo VHD, como fonte de antígenos, associamos a técnica imunoenzimática, com finalidade de facilitar o manuseio e economizar antígeno. O antígeno delta foi obtido a partir do soro de indivíduo na fase aguda de infecção por VHD e o anticorpo a partir de soro de portadores crônicos de VHD. Para a detecção do antígeno foi empregado o método "sanduíche" e para a detecção do anticorpo um ensaio tipo competição. Visando testar a especificidade e a sensibilidade relativas do novo método de detecção de anticorpo, foram feitas comparações do mesmo com radio-imunoensaio comercial (C-RIE, Lab. Abbott) e um radio-imunoensaio desenvolvido na Unité 271-INSERM de Lyon, França (NC-RIA). Soros de 42 pacientes foram testados pelos três métodos, sendo observados resultados discrepantes em apenas 2 casos. Os autores concluem que: 1) soros de pacientes na fase aguda e crônica da infecção pelo VHD podem ser usados como fonte de antígeno e anticorpos em ensaios imunológicos 2) o EIE e o RIE tem especificidade e sensibilidade relativas comparáveis 3) o EIE é de fácil execução, mais barato, não poluente, tem vida útil maior, além de proporcionar economia de antígeno.

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          Most cited references 12

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          Peroxidase-labeled antibody. A new method of conjugation.

           P Nakane,  A. Kawaoi (1974)
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            Epidemiology of HBV-associated delta agent: geographical distribution of anti-delta and prevalence in polytransfused HBsAg carriers.

            The epidemiology of infection with the hepatis-B-virus (HBV)-associated delta agent was assessed from the prevalence of antibody to delta in 1206 HBsAg-seropositive subjects from various parts of the world. Anti-delta was prevalent in unselected HBsAg-positive Italians, whether residents in Italy or elsewhere, and in drug addicts and polytransfused HBsag carriers throughout the world, suggesting that delta-associated infection is spread through contact in Italy and parenterally in other countries. Parenteral transmission of the delta agent was confirmed by a separate survey of the prevalence of anti-delta in 648 polytransfused patients with chronic blood disorders, which showed a higher prevalence of anti-delta in HBsAg-positive haemophiliacs than in the general HBsAg-positive population of Italy, Germany, and the U.S.A. In view of the failure to detect delta in the absence of markers of HBV, the prevalence of anti-delta among polytransfused HBsAg carriers suggests that the delta-associated agent is transmitted by superinfection or coinfection of HBsAg carriers, the HBsAg carrier state possibly providing a rescue function to the superinfecting agent.
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              Diagnostic and prognostic significance of the IgM antibody to the hepatitis delta virus.

              The IgM class antibody to the hepatitis delta virus (HDV) was determined in different clinical categories of hepatitis B surface antigen carriers infected by the HDV (positive in the test for total antibody to HDV). The IgM antibody was found at high titers in each of 70 patients with inflammatory liver disease and at a low titer in one of six patients with inactive cirrhosis; it was not found in eight carriers with normal liver histology. The IgM antibody persisted in high titer over many years in patients with unremitting or progressive liver disease, but declined or disappeared before the homologous IgG antibody in the patients whose disease improved or resolved. Testing for IgM antibody to HDV distinguishes hepatitis B surface antigen carriers who have underlying inflammatory HDV liver disease from those with past HDV infection and provides prognostic information on the course of chronic HDV hepatitis.
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                Author and article information

                Journal
                rimtsp
                Revista do Instituto de Medicina Tropical de São Paulo
                Rev. Inst. Med. trop. S. Paulo
                Instituto de Medicina Tropical (São Paulo, SP, Brazil )
                1678-9946
                December 1987
                : 29
                : 6
                : 388-391
                Affiliations
                São Paulo SP orgnameInstituto Adolfo Lutz orgdiv1Hepatitis Laboratory Brasil
                orgnameUniversidade de São Paulo orgdiv1Faculdade de Medicina
                Lyon orgnameINSERM France
                Article
                S0036-46651987000600010 S0036-4665(87)02900610
                10.1590/S0036-46651987000600010

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 8, Pages: 4
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                Product Information: website
                Categories
                Original Articles

                Hepatitis delta virus, Hepatitis, Enzyme immuno assay

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