Detection of Australia antigen and antibody by means of radioimmunoassay techniques.
01 May 1970-The Journal of Infectious Diseases (Oxford University Press)-Vol. 121, Iss: 5, pp 550-554
TL;DR: It has been shown that the presence of anti-Au does not always prevent the development of hepatitis, and it has been possible to identify Au in some but not all units of blood that have induced hepatitis.
Abstract: Australia antigen (Au) has been identified in the blood of patients with "long-incubation hepatitis" by use of agar-gel diffusion or complement fixation (CF) [1-6]. Characterization of the antigen as a viruslike particle and its localization in the nuclei of liver cells of patients with hepatitis have provided evidence that Au may be the agent of long-incubation hepatitis [7-11]. A precipitating and complement-fixing antibody to Au (anti-Au) has been identified in the serum of patients who have received multiple transfusions but rarely, if ever, in serum from patients convalescing from hepatitis [1-3, 5-12]. It has been shown that the presence of anti-Au does not always prevent the development of hepatitis [13]. Transfusion of blood containing Au may result in hepatitis in the recipient [3, 12]. With agar-gel diffusion, it has been possible to identify Au in some but not all units of blood that have induced
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TL;DR: It is indicated that vertical transmission from carrier mothers frequently occurs, at least in Taiwan, and may partially explain Taiwan's high prevalence of HB5 Ag.
Abstract: To determine the frequency of vertical transmission of hepatitis B antigen (HB5 Ag) from asymptomatic carrier mothers in Taiwan to their offspring, HB5 Ag was sought by radioimmunoassay and complement fixation. Of 158 babies born to carrier mothers, antigenemia developed in 63; 51 of these antigenemic babies had become antigen positive within the six months of life. Three inter-related factors were found to increase the risk that antigenemia would develop in the infant: a high maternal complement-fixation titer for HB5 Ag: presence of HB5 Ag in the baby's umbilical-cord blood: and antigenemia in siblings. In contrast to previous studies, these findings indicate that vertical transmission from carrier mothers frequently occurs, at least in Taiwan, and may partially explain Taiwan's high prevalence of HB5 Ag.
935 citations
Patent•
27 Jan 1972
TL;DR: In this article, a process for the determination of a component of the reaction between a specific binding protein and the substance being specifically bound by such a protein comprising reacting the component to be determined with its binding partner in an insolubilized form, separating the solid phase of a reaction mixture from the liquid phase, reacting with a determined amount of a coupling product of the substance, and finally determining the enzyme activity of the liquid or solid phase obtained.
Abstract: The present invention relates to a process for the determination of a component of the reaction between a specific binding protein and the substance being specifically bound by such a protein comprising reacting the component to be determined with its binding partner in an insolubilized form, separating the solid phase of the reaction mixture from the liquid phase, reacting the solid phase with a determined amount of a coupling product of the substance to be determined with an enzyme, and finally determining the enzyme activity of the liquid or solid phase of the reaction mixture obtained.
851 citations
TL;DR: The hemagglutination test has the sensitivity and rapidity of the best tests available, is simpler to perform, and lends itself to large-scale screening of blood donors.
Abstract: Hemagglutination assays are described for measuring hepatitis-associated Australia antigen and antibody. Red cells coated with isolated antigen, with chromic chloride as a coupling agent, are used for detection of antibodies. Detection of the antigen in serums depends on inhibition of hemagglutination. The test has the sensitivity and rapidity of the best tests available, is simpler to perform, and lends itself to large-scale screening of blood donors.
448 citations
TL;DR: Hepatitis-associated antigen was consistently present in sera from patients with MS-2 strain of serum hepatitis (SH); it was not present in MS-1, infectious hepatitis (IH), and Gamma-globulin consistently neutralized the infectivity of IH (MS-1) serum; in most cases it did not neutralize the infectivities of SH (MS -2) serum.
Abstract: Tests for the presence of Australia or hepatitisassociated antigen (HAA) and antibody (anti-HAA) were performed on more than 25,000 serum specimens from more than 700 patients with viral hepatitis. Hepatitisassociated antigen was consistently present in sera from patients with MS-2 strain of serum hepatitis (SH); it was not present in MS-1, infectious hepatitis (IH). Hepatitis-associated antigen was detected earlier after a parenteral exposure to SH than after an oral exposure. Antigen appeared two weeks to two months before onset of jaundice; it was transient in 65% of patients, but persisted for four months to 13 years in 35% of children. The average incubation period of IH (MS-1) was essentially the same following an oral or parenteral exposure (32 to 33 days); in SH (MS-2) it was 65 days after parenteral exposure and 98 days after oral exposure. Gamma-globulin consistently neutralized the infectivity of IH (MS-1) serum; in most cases it did not neutralize the infectivity of SH (MS-2) serum.
357 citations
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References
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TL;DR: The loss of immunological reactivity at high specific radioactivities or at high levels of chemical substitution with STAI/sup 127/!iodine is demonstrated.
Abstract: A simple and rapid method is presented for the preparation of I/sup 131/- labeled human growth hormone of high specific radioactivity (240-300 mu C/ mu g). Low amounts of carrierfree I/sup 131/ iodide (2 mC) are allowed to react, without prior treatment, with small quantities of protein (5 mu g) in a highyield reaction (approx. 70% transfer of I/sup 131/ to protein). The degree of chemical substitution is minimized (0.5- 1.0 atom of iodine/molecule of protein) by the use of carrier-free I/sup 131/ iodide. The I/sup 131/-labeled hormone (up to 300 mu C/ mu g) contains no detectable degradation products and is immunologically identical with the unlabeled hormone. The loss of immunological reactivity at high specific radioactivities or at high levels of chemical substitution with STAI/sup 127/!iodine is demonstrated. (auth)
10,047 citations
TL;DR: The insulin concentration in plasma has been estimated from the degree of hypoglycemia produced in hypophysectomized, adrenalectomization, alloxan-diabetic rats, and from the increased oxidation of glucose-1-C14 by the rat epididymal fat pad.
Abstract: For years investigators have sought an assay for insulin which would combine virtually absolute specificity with a high degree of sensitivity, sufficiently exquisite for measurement of the minute insulin concentrations usually present in the circulation. Methods in use recently depend on the ability of insulin to exert an effect on the metabolism of glucose in vivo or in excised muscle or adipose tissue. Thus, the insulin concentration in plasma has been estimated: a) from the degree of hypoglycemia produced in hypophysectomized, adrenalectomized, alloxan-diabetic rats (1); b) from the augmentation of glucose uptake by isolated rat hemidiaphragm (2); or c) from the increased oxidation of glucose-1-C14 by the rat epididymal fat pad (3). Since there have been reports indicating the presence, in plasma, of inhibitors of insulin action (4) and of noninsulin substances capable of inducing an insulin-like effect (5,6), these procedures, while yielding interesting information regarding the effects of various plasmas on glucose metabolism in tissues, are of doubtful specificity for the measurement of insulin per se (5).
1,477 citations
812 citations
TL;DR: An antigen that reacted in the immunodiffusion test with serum from multiply transfused patients was detected in the blood during the incubation period prior to the onset of major chemical or clinical abnormalities, suggesting this antigen is specific for serum hepatitis virus.
Abstract: The aim of the following experiments was to provide an objective immunologic criterion for the diagnosis of serum hepatitis, as well as a possible means of screening for carriers of the agent of this disease. An antigen that reacted in the immunodiffusion test with serum from multiply transfused patients was detected in the blood during the incubation period prior to the onset of major chemical or clinical abnormalities. Double blind experiments suggest that this antigen is specific for serum hepatitis virus. Materials and Methods.-Clinical specimens: Sera from cases of transfusion-induced viral hepatitis, which we have collected, were obtained as part of a long-term study involving biweekly follow-up of transfused patients at The New York Hospital. Patients volunteering to participate in this study provided blood samples prior to transfusion and at least biweekly for a period of 6 months or more following transfusion. Test serum: The reference \"antiserum\" used in the majority of the studies to be described, hereinafter referred to as serum S, was obtained from a 24-year-old male patient with hemophilia who has received more than 10,000 units of blood, fresh-frozen plasma, and cryoprecipitate during the course of treatment for bleeding episodes. He has had no episodes of icteric hepatitis, but it was presumed that he had been multiply exposed to the virus or viruses of serum hepatitis. Serum S was chosen for these studies because the patient's multiple exposure was thought to ensure a hyperimmune status. Subsequently, four other sera from multiply transfused patients have been found to react in a manner similar to serum S. For some experiments, the serum was concentrated by ethanol fractionation. To each milliliter of serum to be concentrated, 8 ml of 30% ethanol in 0.1 Ml NaCl, 0.01 M tris(hydroxymethyl)aminomethane (Tris), 0.001 Ml ethylenediaminetetraacetate (EDTA), (pH 7.0 at -7oC) were added. This mixture was held at -70C and lyophilized. The dried globulin fraction was then rehydrated with distilled water to 0.1 the original volume of serum employed. Immunodiffusion technique: Double diffusion in agar gel was done by a micro-Ouchterlony technique.1 Nonspecific precipitation reactions between adjacent wells were eliminated by the use of 0.9% agarose dissolved in a buffer composed of 0.1 M NaCl, 0.01 M Tris (pH 7.6 at 250C), and 0.001 M EDTA containing 1 mg/ml protamine sulfate. Protamine sulfate has been recently suggested as a means of decreasing virus-agar interaction.2 Plates were incubated in a humid atmosphere at room temperature and read daily for 7 days. Strong reactions were evident after overnight incubation, while weaker reactions required 2 or 3 days' incubation and intensified for several days. Clinical chemical methods: Serum glutamic pyruvic transaminase (SGPT) was assayed by a kinetic spectrophotometric method with the Gilford multiple method sample recording spectrophotometer.3 Serum lactic dehydrogenase (LDH) enzymes were assayed by the method of Amador et al.4 with the same instrument. Serum LDH isoenzymes were separated by thin agar gel electrophoresis and quantitated fluorometrically.5 Results.-Demonstration of an antigen appearing in the blood during the incubation period of serum hepatitis: Failure in the past to isolate a causative virus from serum hepatitis could possibly be attributed to the fact that most isolation attempts have been carried out with specimens obtained early in the clinical course of the disease, at what is actually a late stage of the infection due to the
686 citations
"Detection of Australia antigen and ..." refers background in this paper
...Australia antigen (Au) has been identified in the blood of patients with "long-incubation hepatitis" by use of agar-gel diffusion or complement fixation (CF) [1-6]....
[...]
...A precipitating and complement-fixing antibody to Au (anti-Au) has been identified in the serum of patients who have received multiple transfusions but rarely, if ever, in serum from patients convalescing from hepatitis [1-3, 5-12]....
[...]
TL;DR: The precipitin band which forms between the haemophilia antiserum and the serum containing Australia antigen stains faintly with sudan black, indicating that the antigen contains lipid.
Abstract: AUSTRALIA antigen was first identified using an antiserum produced in a transfused patient1,2. The antiserum gave a clear precipitin line in a double diffusion experiment when placed adjacent to the serum from an Australian aborigine. Pending further identification of the antigen, the geographic name “Australian antigen” was given to the reacting material found in the aborigine's serum. Specific antisera against this antigen can be produced by immunizing rabbits with serum containing Australia antigen, and subsequent absorption with serum which does not contain Australia antigen3. The precipitin band which forms between the haemophilia antiserum and the serum containing Australia antigen stains faintly with sudan black, indicating that the antigen contains lipid. It has a specific gravity of less than 1.21 and appears in the first peak in ‘Sephadex G-200’ column chromatography (indicating a high molecular weight)4.
374 citations
"Detection of Australia antigen and ..." refers background in this paper
...Characterization of the antigen as a viruslike particle and its localization in the nuclei of liver cells of patients with hepatitis have provided evidence that Au may be the agent of long-incubation hepatitis [7-11]....
[...]
...A precipitating and complement-fixing antibody to Au (anti-Au) has been identified in the serum of patients who have received multiple transfusions but rarely, if ever, in serum from patients convalescing from hepatitis [1-3, 5-12]....
[...]