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.     

Etiologic structure of viral hepatitis in children based on data from an comprehensive serologic study of hospitalized patients

During a year an overall serological examination of 1,200 children hospitalized with the diagnosis of viral hepatitis was carried out with a view to establish the presence of hepatitis A and hepatitis B markers (anti-HAV IgM, HBsAg, anti-HBs, anti-HBc and anti-HBe IgM) by means of the enzyme immunoassay and radioimmunoassay. The average annual proportion of cases of hepatitis A (61.4%), hepatitis B (20.4%), hepatitis non A, non B (11.2%) and mixed infection (7.0%) among children, as well as seasonal and age-group fluctuations of this proportion, were established. The etiological structure of viral hepatitides, based on commonly accepted clinico-epidemiological diagnostic criteria, was shown to differ from their true etiological structure as determined in accordance with specific serological markers detected in this investigation.     

The prevalence of hepatitis B and C markers among narcotic users

611 patients with acute parenteral virus hepatitis (VH) were studied with a view to find out markers indicating the presence of hepatitis B and hepatitis C virus infection (HB, HC, HB + C, HC + HBsAg). Of these, 166 patients (27.2%) systematically used narcotic drugs intravenously. Essential differences between drug users and VH patients without drug addiction were established regarding the distribution of patients by age and sex, the etiological structure and severity of the disease. Thus, in the group of drug users the prevalence of males, young people (15-29 years of age) and the mixed form of hepatitis B + C was noted. In VH patients using drug the disease took a more severe course than in such patients without drug addiction. The highest proportion of intravenously drug users with a severe and moderate course of the disease was found among patients with HB + C and HB.     

The epidemiological aspects of viral hepatitides in Estonia]

The etiological structure of viral hepatitides among the adult population of Tallinn and the occurrence of markers of hepatitis B and hepatitis C virus infections in medical workers, addict introducing drugs intravenously and hemodialysis patients were studied. Changes in the etiological structure of viral hepatitides were established: they took the form of a decrease in the level of hepatitis A morbidity and the considerable growth of the role of hepatitides B and C, as well as the newly detected circulation hepatitis D virus. About one-third in the structure of morbidity in viral hepatitides were hepatitis cases without markers of hepatitis A, B or C viruses (non-A, non-B, non-C). The highest rates of hepatitis B virus infection (78.9%) and hepatitis C virus infection (82.5%) were detected among drug addicts. Their level of HBsAg was 8.8%. In the group of medical workers, 25% of the examinees, i.e. every fourth person, had markers of hepatitis B virus, while antibodies to hepatitis C virus were detected in 5% of cases. Among hemodialysis patients these rates were 21.4% and 10.7% respectively.     

Seroepidemiological characterization of virus hepatitis in the Republic of Armenia

The survey of the population immunological structure with respect to parenteral hepatitis showed awide circulation of hepatitis B (HB) and hepatitis C (HC) viruses among the adult population of Armenia. During the 5 year period of observation the number of persons having antibodies to HC virus increased 2.7-fold. High occurrence of antibodies to HBsAg of HB virus among the healthy population in 2002 (12.0%) in comparison with 1997 (5.4%) reflected a decreased infection rate with HB virus as well. Antibodies to hepatitis A (HA) virus were isolated, on the average, in 64 % of persons. Simultaneously with a decrease in the proportion of HA cases an increased number of HC patients was registered. No circulation of hepatitis E virus was detected. A high percentage of hepatitis cases of mixed etiology was established, as well as an increased number of combined parenteral hepatitis cases was registered (57.1%).     

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.     

Genetic variability of hepatitis B virus isolates among population of Shuryskarsky area of Yamal-Nenets autonomous region

The purpose of this work was to determine occurrence of serological markers of hepatites B and to describe subtypes of a superficial antigen and genotypes of hepatitis B virus (HBV) isolates among indigenous population of Yamal-Nenets Autonomous Region (YNAR), Russia. METHODS: We investigated 657 serum samples from inhabitants of Shuryskarsky area of YNAR. ELISA method was used to define the hepatitis B markers: HBsAg, anti-HBs (total) and anti-HBc (IgG and IgM). The HBsAg-positive samples were PCR-tested for the presence of HBV DNA. Genotyping of isolates was by sequencing of the Pre-Sl/Pre-82/S region of HBV genome and phylogenetic analysis. Definition of HBsAg subtypes was executed by two methods: ELISA with subtype-specific monoclonal antibodies and S-gene nucleotide sequence analysis. RESULTS: The following occurrence of hepatitis B markers was observed: HBsAg - 3.2%, anti-HBs (total) - 36.2%, anti-HBc IgG - 30.3%, anti-HBc IgM - 1.6%. Frequency of carrying even one of the markers in the observed population was 47.5%. HBV DNA was found in 17 HBsAg-positive samples. Pre-SI, Pre-S2 and S regions sequences were determined for all HBV DNA-positive samples. The phylogenetic analysis showed an accessory of all investigated HBV isolates to genotype D. HBsAg subtypes distribution appeared the following: ayw2 - 23.5%, ayw3 - 70.6%, adw2 - 5.9%. Results of definition of the subtype ELISA method and by the analysis of S gene nucleotide sequences have coincided in 10/11 (90.1%) cases. CONCLUSIONS: The indigenous population of Shuryskarsky area of YNAR belongs to groups with average HBV carrying. Absolute domination of genotype D (subtypes ayw2, ayw3 and adw2) was revealed. High percentage of concurrence of HBsAg subtypes detected by the ELISA method and method of the analysis of S gene primary structure (90%) was observed. Sequencing of HBV S-gene is preferable to define HBsAg subtypes.     

Increased severity and morbidity of acute hepatitis in drug abusers with simultaneously acquired hepatitis B and hepatitis D virus infections.

Hepatitis D virus (delta agent) markers were present in 111 (36%) of 308 intravenous drug abusers who were positive for hepatitis B surface antigen (HBsAg), 52 of these having hepatitis D virus antigenaemia. IgM antibody to hepatitis B core antigen (anti-HBc IgM) was present in 92 out of 95 subjects tested, indicating that hepatitis D virus and hepatitis B virus infections had been acquired simultaneously. Hepatitis D virus markers were present in three out of four patients with fulminant hepatitis, and in 80 of 223 (36%) with mild or moderate hepatitis compared with four of 29 (14%) of those who were asymptomatic. These proportional differences were significant (p less than 0.001). Hepatitis D virus markers were present in twice as many patients positive for anti-HBc IgM requiring admission to hospital with acute hepatitis compared with outpatients attending a drug treatment centre. Tests on one patient showed complete disappearance of HBsAg, but hepatitis D antigen (HDAg or delta antigen) and hepatitis B e antigen (HBeAg) were still present in serum samples. All five patients with chronic active hepatitis had hepatitis D antibody (anti-HD) compared with seven of 24 (29%) with chronic persistent hepatitis (p = 0.008). Blocking anti-HD persisted for long periods after simultaneous infections with hepatitis B virus and hepatitis D virus but at lower titres than in patients with chronic liver disease.     

Morbidity in acute virus hepatitis B on the territory of Kabardin-Balkaria

Morbidity in acute virus hepatitis B (AVHB) in Kabardin-Balkaria during the period of 1992 to 2003 was analyzed. The dynamics of changes in the age groups of AVHB patients, as well as in the structure of the transmission routes of the disease, was analyzed. The level of AVHB morbidity in the Kabardin-Balkar Republic was lower in recent years than the average level of such morbidity in the whole of Russia. The sharply defined irregularity in the territorial distribution of AVHB cases was established. The highest morbidity rates in AVHB were registered in Nalchik, as well as in Chegem and Prokhladnensk regions. The leading role in the formation of the morbidity in acute virus hepatitis B on the territory of Kabardin-Balkaria belongs to Nalchik, where 56.8% of AVHB cases were registered. In the structure of the transmission routes of AVHB the prevalence of artificial paths was noted; among them, the highest proportion belonged to parenteral medical manipulations in outpatient clinics (32.9%). The proportion of AVHB cases associated with the intravenous use of drugs was 6.9%. In the age structure of AVHB patients adults prevailed, and among them the highest number of cases was registered in the age groups of 20 - 29 years and 30 - 39 years. In 2002 the total proportion of AVHB cases among the patients of these age groups reached 68.3%.     

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.     

Profile of acute infectious markers in sporadic hepatitis E

Laboratory diagnosis of acute infection of hepatitis E virus (HEV) is commonly based on the detection of HEV RNA, IgM and/or rising IgG levels. However, the profile of these markers when the patients present have not been well determined. To clarify the extent of misdiagnosed sporadic hepatitis E in the initial laboratory detection, serial sera of 271 sporadic acute hepatitis cases were collected, detected and the dynamics of each acute marker during the illness course were analyzed. 91 confirmed cases of hepatitis E were identified based on the presentation of HEV RNA, IgM or at least 4 fold rising of IgG levels. 21 (23.1%) hepatitis E cases were false negative for the viral RNA and 40 (44.0%) for rising IgG, because occurrence of these markers were confined to acute phase of infection and viremia had already subsided and antibody level peaked when these patients presented. IgM was detected in 82 (90.1%) cases. It is the most prevalent of the three markers, because the antibody persisted until early convalescence. Nine cases negative for IgM were positive for rising IgG and one was also positive for the viral RNA; all of these nine cases showed high avid IgG in their acute phase sera, which indicated re-infection. In summary, it is not practicable to determine the true occurrence of sporadic hepatitis E. Nevertheless, it could be closely approximated by approach using a combination of all three acute markers.     

Isotype, VH genes, and antigen-binding analysis of hybridomas from newborn normal BALB/c mice

In this study we report on the characterization of a panel of 62 hybridomas generated by fusing unstimulated spleen cells from neonatal (less than 24 hr old) normal BALB/c mice with the non-secreting Sp 2/0 cell line. The vast majority (98%) of these hybridomas secreted Ig but only 20% produced IgM. The isotype of the remaining hybridomas was determined as being IgG2b. Interestingly, when splenocytes from 1-day-old mice were stimulated with LPS for 48 h prior to the fusion event, 84% of the hybridomas were secreting IgM. The hybridoma supernatants were screened either by ELISA or RIA for binding reactivity using a panel of 17 Ag, proportionally divided between self and non-self. A binding reactivity could be assigned in 44% of cases. Of these, 29% were monoreactive, i.e., reactivity occurred with one Ag only, while the remaining 15% were multireactive. The majority (21 of 27) of hybridomas with a defined reactivity were directed against self-Ag. These included autologous red blood cells, DNA, histone H1, thyroglobulin, and Ag of the cell surface of T cells. The frequency of utilization of VH genes was determined using DNA probes for eight VH gene families. While all VH gene families appeared to have been used, one, VH 7183, had a slight but significant (p less than 0.02) higher utilization than expected by random expression. The frequency of all the other VH gene families was not significantly different from random utilization. No correlation was found between Ag reactivity in the supernatants and the utilization of a particular VH gene family. These findings indicate that early in the ontogeny the predominant reactivity of B cells is for self-Ag and, unlike what it is commonly believed, the IgM isotype is not dominant within these endogenously activated B cells at this time of ontogeny when genes from all VH families are utilized.     

乙肝大蛋白检测在乙肝抗病毒治疗中价值的研究

目的探讨乙型肝炎病毒大蛋白(HBV-LP)在抗病毒治疗过程中的临床诊断价值。方法选取乙型肝炎患者1 000例为研究对象和100例健康体检者为对照组进行HBV-DNA、HBV-LP和ALT检验;然后筛选志愿者进行拉米夫定抗病毒治疗,在治疗前、治疗中和治疗后分别采集血清标本进行HBV-LP、HBV-DNA、ALT检验。结果在HBVM模式中,HBV-LP和HBV-DNA的阳性率分别为42.64%和43.91%(P>0.05)。HBV-LP阳性标本中HBV-DNA、HBeAg和ALT阳性符合率分别为96.27%、65.01%和98.55%,HBV-DNA和ALT优于HBeAg(P<0.05)。志愿者进行拉米夫定抗病毒治疗中HBV-LP与HBV-DNA均下降,HBV-DNA下降更快。结论 HBV-LP可以检测病毒复制、抗病毒疗效观察和反映肝损伤。     

Detection of Epstein-Barr viral antibodies in patients with acute viral hepatitis

Serum samples obtained from 44 patients with virus A hepatitis, 23 patients with virus B hepatitis, 65 patients with virus non A, non B hepatitis and 100 healthy adults were studied for the presence of Epstein-Barr (EB) virus in the indirect immunofluorescence test. In this work lymphoblastoid cell lines PH3-J-1 and CN37 were used. Among patients with different forms of hepatitis, the statistically significant elevation of the titers of antibodies to EB virus was detected only in the group of patients with virus non A, non B hepatitis, and in 6 cases the etiological role of EB virus was confirmed by serological and hematological methods.     

The detection of viral hepatitis markers in HIV-infected patients

The determination of markers of virus hepatitides B, C and D in 63 registered HIV-infected persons was made. The use of Russian and foreign EIA systems permitted the detection of markers of virus hepatitides 30.1% of HIV-infected persons, including 26.3% of children. Markers of hepatitis B virus were found to occur in children and adults with the same frequency. Out of 65 persons registered in the Center, 3 persons (4.6%) were drug addicts; of these, 2 were found to have antibodies to antigens of hepatitis viruses. Such persons constituted 3.8% of the total number of HIV-infected persons. Among 8 newly detected and registered HIV-infected adults, 2 were found to have antibodies to hepatitis B virus (of these, 1 used drugs intravenously).     

肝炎病毒与EB病毒重叠感染

为探讨肝炎病毒（ＨＶ）与ＥＢ病毒（ＥＢＶ）重叠感染的状况和后果,我们用免疫酶法对１５４例各型病毒性肝炎患者作了ＥＢＶＩｇＡ抗体检测。结果发现,急性肝炎、慢性轻度肝炎、慢性中度肝炎、肝炎肝硬化、慢性重型肝炎和原发性肝癌ＶＧＡ－ＩｇＡ抗体的阳性率分别为２４．０％、３０．０％、５３．３％、６３．３％、４０．０％和７２．７％,与健康人（５．３％）比较,有非常显著升高（Ｐ＜０．０１）;原发性肝癌又较急性肝炎和慢性轻度肝炎高,并有非常显著意义差异（Ｐ＜０．０１）。ＨＢＶ和ＨＡＶ＋ＨＢＶ感染者比较,前者又较后者低（Ｐ＜０．０１）。重叠感染者的临床表现均为“肝炎型”,未见咽炎、腺热、胃肠、肺炎、肾炎、神经等类型。重叠感染者的ＣＤ＋３及ＣＤ＋４Ｔ细胞下降,ＣＤ＋８Ｔ细胞及ＩｇＧ,ＩｇＭ升高,与健康人比较差异非常显著意义（Ｐ＜０．０１）。结果提示：ＨＶ感染,不仅因免疫失调易感ＥＢＶ,又可因重叠感染而进一步使免疫功能失调;对病毒性肝炎的处理应强调免疫调节治疗。     

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.     

Salivary diagnosis of measles: a study of notified cases in the United Kingdom, 1991-3.

OBJECTIVES--To validate a method for salivary diagnosis of measles and to assess the diagnostic accuracy of notified cases of measles. DESIGN--Blood and saliva samples were collected within 90 days of onset of symptoms from patients clinically diagnosed as having measles and tested for specific IgM by antibody capture radioimmunoassay. SETTING--17 districts in England and one in southern Ireland during August 1991 to February 1993. SUBJECTS--236 children and adults with measles notified by a general practitioner. RESULTS--Specific IgM was detected in serum in only 85 (36%) of the 236 cases. In cases associated with outbreaks and tested within six weeks of onset, 53/57 (93%) of samples were IgM positive, thereby confirming the sensitivity of serum IgM detection as a marker of recent infection. The serological confirmation rate was lower in cases with a documented history of vaccination (13/87; 15%) than in those without (70/149; 47%) and varied with age, being lowest in patients under a year, of whom only 4/36 (11%) were confirmed. Measles specific IgM was detected in 71/77 (92%) of adequate saliva samples collected from patients with serum positive for IgM. In cases where measles was not confirmed, 6/101 had rubella specific IgM and 5/132 had human parvovirus B19 specific IgM detected in serum. CONCLUSIONS--The existing national surveillance system for measles, which relies on clinically diagnosed cases, lacks the precision required for effective disease control. Saliva is a valid alternative to serum for IgM detection, and salivary diagnosis could play a major role in achieving measles elimination. Rubella and parvovirus B19 seem to be responsible for a minority of incorrectly diagnosed cases of measles in the United Kingdom and other infectious causes of measles-like illness need to be sought.