Screening for circulating immune complexes in blood donors as an effective method of further HBsAg carriers detection

The loss of counterimmunoelectrophoretic (CIEP) HBsAg reactivity resulting from circulating immune complexes (CIC) formation, observed earlier, prompted us to evaluation the CIC screening in blood donors as an aid in HBsAg detecting. CIC were examined by means of a simple screening modification of the Pegikem test. Among 2.150 normal blood donors there were 21 (0.98%) CIC carriers found, in 13 (61.90%) of them HBsAg was then proved by means of a third generation test. In this population, CIC presence indicated twice as many HBsAg carriers as the CIEP. HBsAg is evidently a very important source of CIC in normal population. The CIC presence is supposed to be a good indicator of HBsAg as well. Therefore, before being accepted as blood donors, CIC carriers should be tested by sensitive test for HBsAg presence.     

High-titered anti-HBs plasma donors--another form of post-hepatitic immunopathy syndrome

In the population of 55 high-titered anti-HBs donors only 23 tolerated plasmapheretic collections without intermittent elevations or ALT activity. In 4 persons a RIA-detected HBsAg circulated along with high-titered anti-HBs. In 73.8% of donors anti-HBs was accompanied by an anti-HBe antibody which also appeared in the HBIG preparation HEPAGA and can perhaps participate on its protective influence. Circulating immune complexes (CIC) were detected in 89.1%. No HBsAg, HBeAg, or albumin were detected in CIC isolated from anti-HBs sera in spite of their content in CIC isolated from HBsAg carriers. Thus, CIC carriers found in normal population with a prevalence of 1.0% can be divided into 0.6% of HHsAg-containing CIC and 0.4% of HBsAg-lacking CIC carriers with anti-HBs attesting the hepatitic origin in a considerable part of them. The continuing production of alienated CIC-forming antigens and a common origin combine these two forms of post-hepatitic development to a syndrome of post-hepatitic immunopathy which seems to be the most frequent source of CIC in a normal population. All the donors and HEPAGA were anti-HBc positive, as well, but this antibody possessed the IgM character only in 4.3% of the donors. Mean serum ferritin levels in the anti-HBs donors were distinctly higher than those found in normal populations of both men and women but the differences were statistically not significant due to high variability.     

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 of antigen-specific determination of HBsAG-containing circulating immune complexes (CIC HBsAG/IgM and CIC HBsAG/IgG) in blood serum of patients with HBV-infection]

A complex enzyme immunoassay (ELISA) has been designed for antigen-specific determination of HBsAg-containing circulating immune complexes (CIC HBsAg/IgM and CIC HBsAg/IgG) in human blood sera in parallel with registration of free HBsAg and specific antibodies to viruses of hepatitis A, B and D. It is shown that effective formation of HBsAg-containing CIC serologically is registered predominantly as a mutually incompatible marker with detection of free HBsAg (in 70-85% of the cases). CIC HBsAg/IgM and CIC HBsAg/IgG may be registered both in parallel and as mutually exclusive markers. Effective formation of HBsAg-containing CIC in the presence of anti-HBsAg occurs in case of a mild course of viral hepatitis of epidemic and sporadic type, while in severe forms of VH-free HBsAg is predominantly detected thus pointing either to ineffective formation of HBsAg-containing CIC or to their continuous registration with demonstration of the effect of delay of witching of anti-HBsM over to anti-HBsG (or CIC HBsAg/IgM to CIC HBsAg/IgG). It was also found that in case of epidemic VH in Tajik SSR (1987) serologically marked as VH both A and B convalescent phase was characterized by parallel disappearance (or lowering of the titer levels) of HBsAg-containing CIC and class M antibodies to both hepatitis A (anti-HAV M) and B (anti-HBcM, anti-HBsM) along with the containing parallel registration of relevant G-antibodies (anti-HAV G/anti-HBcG). This observation requires further studies both in terms of close association of viruses of hepatitides A and B and with regards to possible antigenic mimicry.     

Hepatitis B antigenemia among blood donors: the changing scene.

For the period May 1973 to August 1974 inclusive, the mean prevalence of hepatitis B antigenemia (HBsAg) in all Canadian provinces (per 100,000 population) was, for "first-time" donors, 242 and, for "repeat" donors, 77. A modification of counterimmunoelectrophoresis was used in all 16 regional transfusion centres. The findings confirm the previously noted high prevalence for the Province of Québec and the continuing relatively high prevalence for Canada. The prevalence of HBsAg among donors could be lessened by institution of the following measures: development of a more effective technique for the screening of hepatitis carriers, study of nonparenterally transmitted hepatitis, better reporting of post-transfusion hepatitis, and greater discrimination in prescribing blood and blood components.     

Anti-HBbrno: a new CIEP hepatitis B marker, more sensitive than HBsAg radioimmunoassay.

Evidence is presented for the new HBbrnoAg-Anti-HGbrno system association with the HBsAg. The association is displayed particularly in a prevalence of Ausria detectable HBsAg carriers among the anti-HBbrno carriers, suggesting the anti-HBbrno to be a marker of contact with hepatitis B or, possibly, an indicator of co-existing HBsAg antigenaemia, not detectable by radioimmunoassay. Identity of HBbrnoAg with HBsAg/a, d, y, w, is ruled out, the identity with the other HBV associated antigenic specificities remaining questionable.     

Prevalence and significance of hepatitis B surface antigen in a general hospital.

Over a 6-month period 2025 patients admitted to New Mount Sinai Hospital, Toronto were screened for hepatitis B surface antigen (HBsAg) by counter-immunoelectrophoresis (CIEP) and radioimmunoassay (RIA). CIEP detected 12 HBsAg-positive patients and RIA 16. RIA is therefore the more sensitive test for HBsAg. Of the 16 patients 2 had liver disease previously diagnosed, 3 had malignant disease and 11 were asymptomatic carriers. Of the 11 carriers all were born in countries where the carrier rate is known to be high. Routine screening of hospital patients on admission is of no value in detecting unsuspected liver disease but is of value in detecting asymptomatic carriers, which is of importance for the patient and his family. Routine screening tests for HBsAg in Canadian hospitals that treat many patients born in countries with a known high HBsAg prevalence is recommended. Routine screening is also recommended in all hospitals in Mediterranean and Asian countries.     

Genotype and subtype analyses of hepatitis B virus (HBV) and possible co-infection of HBV and hepatitis C virus (HCV) or hepatitis D virus (HDV) in blood donors, patients with chronic liver disease and patients on hemodialysis in Surabaya, Indonesia

Four subtypes (adw, adr, ayw, and ayr ) and eight genotypes (A to H) of the hepatitis B virus (HBV) have been identified. They appear to be associated with particular geographic distribution, ethnicity, and possibly clinical outcomes. In this study, hepatitis B surface antigen (HBsAg) subtyping and HBV genotyping were carried out on sera obtained from HBsAg-positive HBV carriers, including healthy blood donors; patients with acute hepatitis, chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma; and patients on hemodialysis all located in Surabaya, Indonesia. We report here that all HBV isolates tested in Surabaya belonged to genotype B, with more than 90% of them being classified into subtype adw. Our results also revealed that prevalence of hepatitis C virus (HCV) co-infection among HBV carriers in Surabaya was approximately 10% for healthy blood donors and patients with chronic liver disease, and approximately 60% for patients on maintenance hemodialysis. Interestingly, HBsAg titers were lower in HBV carriers with HCV co-infection than in those without HCV co-infection. We also found that prevalence of hepatitis D virus (HDV) co-infection was < 0.5% among HBV carriers in Surabaya.     

Hepatitis B virus DNA and e antigen in serum from blood donors in the United Kingdom positive for hepatitis B surface antigen.

Serum samples from 214 blood donors in the United Kingdom who were carriers of hepatitis B surface antigen (HBsAg) were examined for hepatitis B virus deoxyribonucleic acid (DNA) by DNA:DNA hybridisation and for hepatitis B e antigen (HBeAg) and its antibody. One fifth of the donors carried infectious virus in their circulation. The presence of hepatitis B virus DNA correlated well with that of HBeAg, although hepatitis B virus DNA was found in five serum samples that were negative for HBeAg. It is concluded that analysis of serum samples for hepatitis B virus DNA by hybridisation should be the method of choice for determining whether carriers of HBsAg are infectious.     

New strategies for blood donor screening for hepatitis B virus: nucleic acid testing versus immunoassay methods

Serologic testing for hepatitis B virus (HBV) surface antigen (HBsAg) and antibody to HBV core antigen (anti-HBc) has historically been the foundation of blood screening, while HBV nucleic acid testing (NAT) was recently developed to detect HBsAg-negative, anti-HBc-negative blood units donated during early acute infection. Comparison data on seroconversion panels using HBsAg assays of varying sensitivities and pooled- or single-sample NAT, along with viral load estimates corresponding to HBsAg assay detection limits, have provided information on the theoretical benefits of NAT relative to HBsAg. Model-derived estimates have generally been predictive of the yields of DNA-positive, HBsAg-negative window period blood units detected in a number of studies from Europe, Japan, and the US. Studies indicate that the added benefit of pooled-sample NAT is relatively small in areas of low endemicity, with greater yields in areas highly endemic for HBV. Single-sample NAT would offer more significant early window period closure and could prevent a moderate number of residual HBV transmissions not detected by HBsAg assays; however, no fully automated single-sample HBV NAT systems are currently available.Even single-sample HBV NAT may not substitute for anti-HBc screening, as indicated by studies of donors with isolated anti-HBc who have extremely low DNA levels undetectable by standard single-sample NAT and who have been associated with transfusion-transmitted HBV. Moreover, HBsAg testing may still be needed even in the setting of combined anti-HBc and NAT screening. HBsAg-positive units from donors in the chronic stage of infection may contain very low or intermittently detectable DNA levels that single-sample NAT would miss. Although such donors are usually anti-HBc reactive and would be interdicted by anti-HBc screening, some lack anti-HBc. Extensive parallel testing will be needed to determine whether single-sample NAT in combination with anti-HBc might be sufficient to detect all the infectious donors currently interdicted by HBsAg testing. In countries that do not screen for anti-HBc, HBsAg testing would be the only means of detecting donations from chronically infected individuals with low/intermittently detectable DNA, since even single-donor NAT would not identify these potentially infectious blood units. In the future, the current fully automated HBsAg assays may incorporate significant sensitivity improvements, and automated single-sample HBV NAT may become a reality. Each country will need to develop its blood screening strategy based on HBV endemicity, yields of infectious units detected by different serologic/NAT screening methods, and cost effectiveness of test methods in ensuring blood safety.     

Characteristics of the formation of the HBsAg carrier state among pregnant women and newborn infants and their role in the spread of hepatitis B

The authors analyze the incidence rate of HBsAg carriership among 8, 120 pregnant women and 261 newborn infants at different periods after birth. The levels of HBsAg carriership among pregnant women and the members of their families, as well as among the personnel of maternity clinics and blood donors, have been established. The rate and time of the detection of HBsAg in infants born to mothers found to be HBsAg carriers have been determined. Measures for the prophylaxis of hepatitis B are discussed with due regard to the specific epidemiological features of the spread of HBsAg carriership, established in this study, and to the presence of antibodies to HBsAg among the above-mentioned groups of the population.     

Plasma parathyroid hormone during acute volume expansion in the rat

Various studies have suggested the possibility that volume expansion may increase parathyroid hormone (PTH) secretion. PTH appears to have renal effects consistent with the actions of a natriuretic and diuretic and the possibility exists that PTH may play a physiological role in volume homeostasis. The present studies were designed to examine whether PTH levels in plasma from rats was influenced by acute volume expansion and whether such effects were independent of alterations in plasma ionized calcium concentration. Volume expansion with calcium-free bicarbonate Ringers (10% of body weight, IV) led to a drop in plasma ionized calcium from 1.08 to 0.92 mMol/l (p less than 0.01) while plasma PTH concentration was increased from 67.2 to 114.2 pMol/l. Volume expansion with bicarbonate Ringers solution (also 10% of body wt, IV) which contained 1.8 mM CaCl2 was not associated with any significant change in either plasma ionized calcium or plasma PTH concentration. However, measurements of blood packed cell volume (PCV) revealed that infusion resulted in a drop in PCV from 49.7 to 41.1% (p less than 0.01). This represents a dilution of plasma of approximately 42%. The absence of any drop in plasma PTH during isocalcemic volume expansion suggests an underlying stimulus to PTH secretion during volume expansion independent of plasma ionized calcium levels.     

Liver enzyme concentrations as measure of possible infectivity in chronic asymptomatic carriers of hepatitis B.

Fifty-two British-born blood donors who were chronic carriers of hepatitis B surface antigen (HBsAg) were tested for the presence of hepatitis B e antigen (HBeAg) and antibody to HBeAg by an immunoradiometric assay. The presence of HBeAg was closely associated with a slight rise in serum liver enzyme concentrations, a high HBsAg titre, and male sex. We suggest that the finding of persistently raised serum liver enzyme concentrations in an asymptomatic HBsAg carrier might be useful as a likely indicator of HBeAg and high infectivity.     

婴儿围产期感染乙型肝炎病毒后各项血清学指标的动态变化

本文观察了102名新生儿出生后至18月龄的HBV血清学指标的动态变化,婴儿分成乙型肝炎疫苗按种组(63人)和对照组(39人), 观察期间HBsAg始终阴性的70名婴儿,出生后6、12和18月龄的抗-HBc阳性率依次为90％、30％和4.3％;而HBsAg阳转的27名婴儿,18月龄时抗-HBc全都阳性,但仅有6名婴儿在6月龄时测出IgM抗-HBc,疫苗接种组婴儿出生后1、3、6、12和18月龄的抗-HBs阳性率,依次为28.6％、76.2％、77。8％、82.5％和82.5％;对照组婴儿18月龄时抗-HBs阳性率仅为12.8％。     

Success of a program of routine prenatal screening for hepatitis B surface antigen: the first 2 years.

Prenatal screening for hepatitis B surface antigen (HBsAg) restricted to women with defined risk factors for chronic hepatitis B virus (HBV) infection fails to identify many carriers. A centralized program of routine HBsAg screening for all pregnant women in Alberta was introduced in 1985. We collected and analysed data for the first 2 years of the program in Edmonton to determine the frequency of risk factors for HBsAg positivity, the proportion of multiparous HBsAg-positive women not identified in previous pregnancies, the efficiency and cost-effectiveness of providing immunoprophylaxis to infants at risk of HBV infection and the degree of success in inducing adequate protection. A total of 149 women (158 pregnancies) were found to be HBsAg positive. Risk factors were readily ascertainable for 85% of the women; the remaining 15% would not have been identified through risk-selective screening. The most common risk factors were Oriental ethnic origin, history of hepatitis, jaundice or multiple transfusions of blood or blood products, and occupational exposure to blood. Although 86% of the multiparous HBsAg-positive women had risk factors, only 7% had been identified in previous pregnancies. The Alberta program appears to be cost-effective. We conclude that only routine prenatal screening will identify all infants at risk of perinatal HBV infection and that a comprehensive public health program involving central laboratories, private physicians and public health staff can be highly effective and efficient in protecting infants against hepatitis B.     

Algorithm of serologic screening and assessment of prevalence of serologically meaningful mutations of HBsAg in hepatitis B virus carriers

Algorithm of serologic screening for HBsAg-mutants in hepatitis B virus (HBV) carriers with high level of HBsAg was developed which is based on the detection of defects of interactions of serum HBsAg with monoclonal anti-HBs realizing as a decrease of ELISA sensitivity in 10 times or more during serial 10-fold dilutions. During 1st stage commercial test-systems based on monoclonal antibodies was used to select serum samples with discrepancy of test results. During 2nd stage HBsAg contained in selected sera was analyzed by the panel of monoclonal and polyclonal anti-HBs conjugates using decrease in ELISA sensitivity as a criterion. Serum samples from 2510 chronic carriers of HBV with high level of HBsAg were studied. 19 samples with discrepant results were found. Subsequent characterization of HBsAg with panel of 11 monoclonal and 1 polyclonal conjugates allowed to distinguish groups of sera with specific serologic "portraits". Atypical features of HBsAg were confirmed by genotyping 9 of 19 samples. Analysis of primary nucleotide sequence revealed serologically meaningful mutations in S-gene of HBV in all 9 isolates: 3 of them contained substitution mutation G145R, 5--S143L, and one--T143M. Distribution of mutations in HBsAg corresponded with specific serologic "portraits". Prevalence of HBsAg mutations in HBV carriers with high level of HBsAg was assessed for the first time: prevalence of G145R, S143L/T143M mutations, and all serologically atypical variants was 0.12%, 0.24%, and 0.76% respectively. Developed algorithm was proposed for epidemiologic monitoring of HBsAg-mutants of HBVand control of diagnostic test-systems.     

Prevalence of intestinal parasitic infestation,salmonellosis, brucellosis,tuberculosis, and hepatitis B among immigrant children in Glasgow.

Two hundred Asian and 100 each of African, Chinese, and Scottish children were screened for intestinal parasitic infestations, salmonellosis, brucellosis, hepatitis B antigen (HBsAg), and tuberculosis. There was a fairly high incidence of Giardia lamblia among Asian and Scottish children and of Trichuris trichiura among the Chinese. Hookworm ova were seen only in Africa children. There were no chronic carriers of Salmonella or Brucella, and no one was suffering from salmonellosis or brucellosis. Tuberculin sensitivity was found in only 4% of immigrant and 1% of Scottish children: the difference was small and neither figure suggests a continuing high incidence of tuberculosis in Glasgow. Only seven immigrant children were found to be HBsAg carriers. Among the families of these carriers there was a high incidence (84%) of HBsAg or antibody (HBsAb). The survey shows that immigrant children in Glasgow do not constitute a health hazard to the indigenous population. Moreover, severe overcrowding is not a prominent feature among the immigrant families in Glasgow but is greatest among the local Scots.     

Prevalence of hepatitis B viral markers in Vietnamese blood donors

Serum samples were assayed using radioimmunoassay in 573 Vietnamese blood donors living in Hano? (North Viet Nam). 66 (11.5%) subjects were HBsAg-positive. Of these 66 HBsAg carriers, 17 (25,8%) were positive for hepatitis B e antigen (HBeAg) and 43 (65.1%) for antibody to HBeAg (anti-HBe). 22 (3.8%) subjects were positive for antibody to hepatitis B core antigen (anti-HBc) alone. 402 (70.2%) subjects were positive for antibody to HBsAg (anti-HBs). This anti-HBs percentage increased with age. Only 83 (14.5%) subjects were negative for all hepatitis B viral (HBV) markers. This no HBV markers percentage decreased with age. The chi 2 test showed a non significant difference for frequencies of HBsAg, anti-HBc alone, anti-HBs but a significant one for frequencies of no HBV markers in men and women.     

Blood transfusion and hepatitis: still a threat?

The incidence of post-transfusion hepatitis (PTH) in recipients of blood products is reviewed. PTH was observed in 10%-12% of recipients of blood products in the United States, 2%-4% in northern Europe and 15%-20% in southern Europe. All studies indicate that 80%-90% of all PTH cases are attributed to non-A/non-B. At least 40% of the patients with PTH non-A/non-B will develop chronic hepatitis or cirrhosis. No specific tests for the detection of the non-A/non-B agent(s) exist. However, several independent studies indicate that part of the donors carrying the infectious non-A/non-B agent have increased levels of alanine amino transferase (ALT). When donors are excluded with elevated ALT values, it is estimated that about 30% of the PTH non-A/non-B cases would be prevented. Some studies indicate that anti-hepatitis B core (anti-HBc) positive donors may carry an increased risk to transmit the non-A/non-B agent, but more recent studies do not confirm this. There is hope that a specific non-A/non-B test will be developed soon.     

IgM antibody against hepatitis B core antigen (IgM anti-HBc): diagnostic and prognostic significance in acute HBsAg positive hepatitis.

IgM antibody against hepatitis B core antigen (IgM anti-HBc), a marker of recent hepatitis B virus infection, was sought by radioimmunoassay in sera diluted 1/4000 from 376 patients presenting to four centres in Italy with acute, apparently type B hepatitis (hepatitis B surface antigen (HBsAg) positive). In 320 patients (85%) a positive IgM anti-HBc test result confirmed that hepatitis was due to primary infection with hepatitis B virus. In the remaining 56 patients absence of the IgM marker indicated that they were previously unrecognised long term carriers of HBsAg. Further serum analysis often showed delta infection and occasionally hepatitis A or cytomegalovirus infection as the true cause of their illness. After six to eight months circulating HBsAg persisted in 38 of 45 patients (84%) without IgM anti-HBc but in only six of 150 patients (4%) with the IgM antibody (p less than 0.0001). A negative IgM anti-HBc test result in patients with acute HBsAg positive hepatitis points to a factor other than hepatitis B virus as the cause of the liver damage and predicts the carriage of HBsAg.