The danger of hepatitis B virus infection from transfusions of plasma in which the HBs-antigen was detected retrospectively by highly sensitive methods

The prospective dynamic laboratory and clinical study of premature children was carried out: 55 children who received plasma transfusion during the first weeks of their life and were retrospectively (i. e. after plasma transfusion) found to have HBsAg detected by the passive hemagglutination (PHA) test, the enzyme immunoassay (EIA) and the radioimmunoassay (RIA) and 127 children who received the transfusion of plasma found to be HBsAg-negative according to the results of EIA and RIA. As revealed in this study, the risk of hepatitis B virus infection in such children after the transfusion of plasma containing HBsAg at low concentrations, determined only in the PHA test or in EIA and RIA, was, respectively, 7.5 and 6.3 times greater than in children receiving plasma found to be HBsAg-negative according to the results of EIA and RIA. The results of this investigation showed the tendency towards a decrease not only in the total contamination of plasma recipients with hepatitis B virus, but also in morbidity rate in icteric forms of acute posttransfusion hepatitis B, depending on the concentration of HBsAg in plasma used for transfusion.     

Detection of HBsAg in the familial environment of children with viral hepatitis B

A total of 231 persons from the families of 62 children hospitalized in connection with viral hepatitis B were examined for the presence of HBsAg in their blood over a period of 3 years. Simultaneously with countercurrent electrophoresis (CIE) and the gel precipitation (GP) test, the passive hemagglutination (PHA) test and the GP test with sandwich treatment were used in this work. The presence of HBs-antigenemia in the members of the families of children with viral hepatitis confirmed by laboratory methods were found to occur 6.7 times more frequently than in the families of children with HBsAg-negative hepatitis. The use of the PHA test and the GP test with sandwich treatment increased the frequency of the detection of HBsAg 2.5 times in comparison with CIE and the GP test. The data indicating the possibility of children being infected through everyday contacts in families with cases of HBs-antigenemia among their members are presented, but further studies are necessary to make the final decision on this problem.     

The results of a controlled field trial of a dried erythrocytic immunoglobulin diagnostic agent for detecting the hepatitis A virus antigen]

The results of the controlled field trial of lyophilized erythrocytic immunoglobulin diagnosticum for the detection of hepatitis A virus antigen in the urine and feces of patients are presented. This diagnosticum was used for the study of urine and fecal samples from 225 patients (of these, 176 had hepatitis A) and 54 healthy persons in the passive hemagglutination (PHA) test. Their blood sera were studied in the PHA test (to detect HBsAg) and the radioimmunoassay (to detect anti-HAV IgM). The immunoglobulin diagnosticum under study was found to be nonspecific and faintly sensitive and, therefore, unsuitable for use in medical practice.     

Epidemiological characteristics and the importance of carriers of the surface antigen of the hepatitis B virus (HBsAg)

The occurrence of HBsAg carriership in Leningrad has been found to be 1.4% according to the results of countercurrent immunoelectroosmophoresis (CIEO) and 2.1% according to the results of the passive hemagglutination (PHA) test. In children HBsAg occurs with higher frequency: 1.9% according to the results of CIEO and 3.4% according to the results of the PHA test. The latter test reveals HBsAg carriers more completely, especially in women who have usually less pronounced antigenemia than men. Most of chronic HBsAg carriers are patients with chronic forms of hepatitis B (chronic active hepatitis and chronic persistent hepatitis); frequently they become the source of infection among their relatives under the conditions of family contacts. A complex of antiepidemic measures is necessary in the foci of chronic HBsAg carriership.     

The territorial and age-related characteristics of the distribution of patients with non-A, non-B viral hepatitis infected via the fecal-oral mechanism in foci of elevated viral hepatitis morbidity in the Uzbek SSR

The study of patients from 10 foci of acute viral hepatitides for the presence of HBsAg (in the passive reverse hemagglutination test) and anti-hepatitis A virus IgM (in the radioimmunoassay) has shown high frequency and variability in the spread of hepatitis non-A, non-B, the prevalence of adults aged 20-29 years and children aged 2-4 years among persons involved into the epidemic process and the tendency towards an increase in the proportion of hepatitis non-A, non-B in the total number of cases of viral hepatitides in the republic.     

Comparative evaluation of methods for detecting HBsAg in sera

The results of the analysis of 1209 serum samples, made with a view to detecting those containing HBsAg, are presented. This analysis was made by the radioimmunoassay (RIA) on a polyethylene film, by the standard RIA technique with the use of a diagnostic kit obtained from Abbott Laboratories (USA) and by the passive hemagglutination (PHA) test. The RIA film technique was found to have the sensitivity of about 2 ng/ml HBsAg, which is similar to the sensitivity of the kit from Abbott Laboratories and exceeds the sensitivity of the PHA test approximately 50-fold. The percentage of detected HBsAg-positive sera, yielded by analysis with the use of the RIA film technique and the standard RIA technique, is the same. The RIA technique make it possible to detect more positive sera than the PHA test by about 2.5%.     

Frequency of finding hepatitis B viral markers in young children born of mothers carrying the HBs-antigen

The present communication deals with the results of investigations on the frequency of HBsAg-carriership among expectant mothers in Fergana (the Uzbek SSR) and on the risk for their children to be infected with hepatitis B virus. To detect the markers of hepatitis B virus, the passive hemagglutination test, the enzyme immunoassay, and the radioimmunoassay were used. The incidence rate of HBsAg-carriership among expectant mothers was rather high (5.1 +/- 0.52%) in comparison with the control group (3.5 +/- 0.70%). The detection of HBsAg in children 3 months after their birth, i.e. at the term corresponding to the possible incubation period of hepatitis B, suggested that the children were infected at birth. As noted in the course of this investigation, the risk for a newborn to be infected was directly related to the titer of HBsAg, as well as to the presence of HBeAg, in the blood of the carrier mother. A high detection rate of HBsAg was registered among children born to HBsAg-carrier mothers, which makes it possible to consider these children as a high risk group with respect to hepatitis B virus infection and necessitates the development of the system of antiepidemic and prophylactic measures for preventing newborns to be infected with hepatitis B.     

Frequency of detecting viral hepatitis B markers in the patients of a hemodialysis ward

The data, obtained as the result of the examination of 22 patients with chronic renal insufficiency and analysis of 105 samples of transfusion blood at the department of chronic hemodialysis, are presented. To detect the markers of hepatitis B (HBsAg, anti-HBs, anti-HBc), a complex of biochemical investigations was carried out with the use of counter-current immunoelectrophoresis, the passive hemagglutination test, enzyme immunoassay, and solid-phase radioimmunoassay. The markers of hepatitis B were detected in 72.6% of patients with chronic renal insufficiency and in 21% of healthy persons. Changes in the activity of biochemical characteristics of hepatic samples were detected only in one patient. In no case clinical symptoms of the disease were observed. Out of 105 samples of transfusion blood, 9.5% contained HBsAg. The results of our investigations indicate that the markers of hepatitis B are widely spread among patients with chronic renal insufficiency, which makes it possible to consider them as a "high risk group" with respect to hepatitis B infection. To decrease the risk of hepatitis B among patients with chronic renal insufficiency, it is very important that highly sensitive tests be introduced into practice for the selection of donors and the detection of patients with the asymptomatic forms of hepatitis B and carriers at the department of chronic hemodialysis.     

Etiological structure of viral hepatitis in the area about one of the cities in Gorki Province

The etiological structure of viral hepatitides (VH) in one of the towns of the Gorky region was studied with the use of specific methods for diagnosing hepatitis A (detection of IgM to hepatitis A virus) and hepatitis B (detection of HBsAg in the passive hemagglutination test). The study revealed that hepatitis A was the major nosological from in the structure of VH among children and adults in the area under survey, which was documented by the detection of IgM to hepatitis A virus. The form, second in importance among VH cases, was hepatitis B. The ratio of these two forms of VH was determined by the epidemiological situation in the area. The proportion of hepatitis B cases increased at the period between epidemics. Nondifferentiated hepatitis constituted 6.8% of all cases of sporadic hepatitis among adults. In 90% of cases clinical diagnosis coincided with the serological one.     

Results of the examination of patients with viral hepatitis for HBsAg in Kashka-Darya Province, Uzbek SSR

The practical use of the reverse passive hemagglutination test for the examination of viral hepatitis patients revealed the presence of HBsAg, on the average, in 28.4 +/- 0.95% of cases. This antigen occurred most frequently among the patients aged up to 1 year (42.8 +/- 4.18%), as well as in the second quarter of the year (33.3 +/- 2.3%). The proportion of patients with hepatitis B in the total number of patients with viral hepatitis was 20%.     

The risk of infecting premature newborn infants with the HB virus in transfusions with plasma HBsAg-negative by counterimmunoelectrophoresis

The prospective dynamic study of 229 premature infants with the use of highly sensitive methods, such as the passive hemagglutination test and the enzyme immunoassay, for the detection of HBsAg and anti-HBs antibodies. Of these, 182 infants received plasma (blood) transfusions in the first days of their life. Plasma (blood) used for transfusions was known to contain no HBsAg in accordance with the results of countercurrent immunoelectrophoresis. 47 infants had no plasma (blood) transfusions. In the group of 182 plasma recipients the presence of hepatitis B virus infection was detected in 17.0% of infants, the ratio of icteric, nonicteric and inapparent forms being 1:4.2:2. In the group of 47 infants no case of hepatitis B virus infection was observed.     

Determination of antibody to hepatitis B core antigen by means of immune adherence hemagglutination.

A simple and rapid method utilizing immune adherence hemagglutination has been developed for the detection of antibodies to hepatitis B core antigen (anti-HBc). Hepatitis B core antigen (HBcAG) was prepared from Dane particles that had been isolated from plasma of asymptomatic antigen carriers. The method was specific and about 10 times more sensitive than the conventional complement-fixation method. A total of 215 serum samples obtained from healthy blood donors were surveyed for HBsAG and anti-HEc, as well as for hepatitis B surface antigen (HBsAg) and antibody to HBsAG (anti-HBs). Anti-HBc was found in 36 serum samples, at a prevalence rate higher than that of anti-HBs (31/215)...     

Screening of Danish blood donors for hepatitis B surface antigen using a third generation technique.

The profit to be gained by testing Danish blood donors for hepatitis B surface antigen (HBsAg) with a third generation technique instead of the currently used immunoelectrophoresis was investigated by additional screening of 48 750 blood units by radioimmunoassay three weeks after donation. Twenty nine units were positive for HBsAg on radioimmunoassay (0.059%). Only six of these were found by immunoelectrophoresis (0.012%). Most of the 23 donors positive on radioimmunoassay and negative on immunoelectrophoresis were healthy carriers of HBsAg (20) or had asymptomatic chronic liver disease (two). One donor had acute hepatitis B. Fifteen of the 23 blood units were transfused. The 15 recipients were monitored biochemically and serologically for up to nine months. One recipient developed fulminant hepatitis B, three developed acute hepatitis B, and one became a healthy carrier of HBsAg. All these patients had received blood from healthy carriers of HBsAg. Two recipients were immunised against HBsAg, and in one patient no seroconversion was observed. The remaining recipients died soon after transfusion or were protected by antibodies to HBsAg that had been present before the transfusion. Testing of Danish blood donors using a third generation technique identified a substantial number of donors positive for HBsAg overlooked by immunoelectrophoresis. Most of these donors were healthy carriers of HBsAg. Blood taken from such carriers is highly infectious when transfused, probably because of the large amount of material transmitted.     

Immunoenzyme analysis for determining class G and M antibodies to HBsAg

A scheme of the purification of hepatitis B virus surface antigen (HBsAg) as applied to the enzyme immunoassay (EIA) for the detection of antibodies to HBsAg is described. An indirect EIA technique for the detection of IgG and IgM antibodies to HBsAg has been developed and the diagnostic assay system based on the use of immunoreagents and solid-phase carriers produced in the USSR has been obtained. The sensitivity of the indirect EIA technique in the detection of IgG antibodies to HBsAg exceeds that of double immunodiffusion in gel used for this purpose 2,500- to 5,000-fold. The study has shown the possibility of using the indirect EIA technique for the detection of antibodies to HBsAg, both free and bound in immune complexes, of detecting antibodies to HBsAg in patients with acute and chronic viral hepatitis B, as well as of simultaneous detection of IgG and IgM antibodies to HBsAg without pseudonegative results.     

The combination of pneumocystosis with the hepatitis B virus carrier state in cancer patients]

In an oncological hospital the occurrence of the combined infection of patients with Pneumocystis carinii and hepatitis B virus was determined. Parasitological investigations for the presence of pneumocystosis were carried out by means of light and luminescent microscopy. Serological investigations for hepatitis B were made in the passive hemagglutination test. The examination of 171 patients with malignant tumors of the head and the neck and 47 patients with benign tumors revealed the presence of P. carinii in 45 patients (26.3%) and 4 patients (8.5%) respectively. Among the patients with malignant tumors, P. carinii and HBsAg were detected simultaneously in 9 patients, i. e. in 20% of all patients infected with P. carinii.     

A trial application of the latex test for evaluation of antibody level in rabbit immune sera]

A latex test was elaborated which served for evaluation of quality of rabbit immune sera for antigen 0 of selected Gram-negative bacteria. Sensitivity and specificity of this test in comparison with passive hemagglutination and immunoenzymatic DOT-ELISA reactions was evaluated. These studies were performed on immune sera for antigen O of Salmonella groups B, C1, C2, D and E, Yersinia pseudotuberculosis and in antigen preparations for above listed microorganisms both in homologous and heterologous systems. It was found that sensitivity of the latex test is 9 to 160 times lower than that of passive hemagglutination and 7 to 307 lower than for DOT-ELISA. Sensitivity of the latex test and passive hemagglutination reaction was evaluated on the basis of results of cross reaction between studied antigens and unabsorbed rabbit sera, establishing so called sensitivity indexes, which were informing how many times heterologous titer is lower than homologous titer. So evaluated sensitivity of the latex test was close to sensitivity of the passive hemagglutination reaction. It was found that slide latex test is characterized by satisfactory sensitivity and good sensitiveness and may be used for evaluation of antibody level 0 antigens of Salmonella and Yersinia. The value of this test is characterized by high repeatability of results, as well as low work and time-consuming.     

Sensitivity and specificity of Czechoslovak ELISA HBsAg Sevac tests: comparison with other third-generation tests and counter-immunoelectrophoresis

Two variants of sandwich-type ELISA (Enzyme Linked Immunosorbent Assay) kits for HBsAg detection (Sevatest ELISA HBsAg Macro I and Sevatest ELISA HBsAg Micro I) in human sera and plasmas were developed. As the solid phase, the ELISA Macro kit and ELISA Micro kit make use of polystyrene microtubes, and polystyrene microtitration plates, respectively, of Czechoslovak production (Koh-i-noor, Dalecín). Capture anti HBs antibody for adsorption to solid phase and rabbit anti HBs antibody for labelling with horse-radish peroxidase were prepared for both tests. The sensitivity of both ELISA kits for HBsAg, equal to approx. 2 ng/ml, was determined by titrating six selected HBsAg-positive sera and the WHO Agk 76 panel of HBsAg-positive sera and the results were compared with those obtained by ELISA, RIA (Radioimmunoassay) and RPHA (Reverse passive hemagglutination) kits of different producers and by counter-immunoelectrophoresis (CIEP). The sensitivity of the new ELISA kits was comparable to that of other producers' ELISA kits, higher than that of RPHA kits and only a little lower than that of RIA kits. A set of sera of patients hospitalised with different diagnoses was tested for HBsAg. The detection rate by ELISA Macro kit 2.8 and 1.5 times higher than by CIEP and RPHA (Raphadex B), respectively, and 1.1 time lower than by RIA (Austria II).     

Characteristics of the distribution of the markers of hepatitis B vir us infection among the healthy population of the Tadzhik SSR and Azerbaijan SSR

A total of 708 healthy persons in Tajikistan and 576 healthy persons in Azerbaijan, these groups comprising persons of both sexes and different age groups, were examined by the method of double gel immunodiffusion (the gel precipitation test) and by the passive hemagglutination test for the presence of the markers of hepatitis B virus (HBV) infection (HBsAg and HBeAg) and antibodies to them. This investigation showed that, in accordance with the level of hepatitis B morbidity, HBsAg was significantly more often detected among the population in Tajikistan (7.2%) than in Azerbaijan (2.8%). In both republics HBV carriers occurred most frequently among children aged 1-4 years (4.0% in Azerbaijan and 13.9% in Tajikistan), and among men more frequently than among women. In accordance with different intensity of the spread of HBV infection in the territories under comparison, differences in the age structure of the immune population were noted: in Tajikistan the formation of the immune layer occurred most frequently among younger age groups and in Azerbaijan, among senior adult age groups. The presence of a considerable percentage of persons with HBe-antigenemia (14.3-14.9% as determined by the gel precipitation test) among HBV carriers, observed in Tajikistan and in Azerbaijan, indicates that some of them have undetected chronic hepatitis B.     

Escherichia-erythrocyte diagnostic agents

The conditions of a simple and practicable method for the preparation of effective antigenic nonprotein diagnosticums on the basis of water-phenol extracts of 23 Escherichia species have been developed. The method consists in heating the mixture of erythrocytes and the antigen in a boiling water bath for 60 minutes. The diagnosticums thus obtained are 16-30 times more sensitive in the passive hemagglutination test and 4-6 times more sensitive in the passive hemagglutination inhibition test than diagnosticums prepared with the use of tannin, rivanol, as well as by the common method for the preparation of nonprotein antigens. The minimum concentration of Escherichia cells detected in the passive hemagglutination inhibition test is 0.8-1.2 million cells/ml.     

Application of the paired label radioantibody technique to detection and subtyping of hepatitis B surface antigen.

A sensitive radioimmunoassay has been developed for the detection and subtyping, in regard to w and r specificities, of hepatitis B surface antigens (HBsAg). The binding of monospecific rabbit antibodies directed to either w or r determinant, labeled with 131I and 125I, respectively, by HBsAg was determined in the same experiment, taking advantage of the fact that these specificities are in general, expressed mutually exclusively. HBsAg in the test sample were bound to solid phase guinea pig anti-HBsAg antibody tube before the test. The ratio of percentage uptake of 125I anti-r to that of 131I anti-w for 28 normal sera without antigen was 0.69 0.18. The ratio for seven sera containing HBsAg with w determinant was less than 0.1, and that for 34 with r determinant was more than 10; 100-fold difference was noted between values of w and r antigens.