The prevalence of molecular and immunologic infective markers of hepatitis viruses in patients with hematological malignancies

Acute and chronic viral hepatitis infections are corresponding to increase the risk of different types of hematological malignancies especially with leukemia. In this study the serological and molecular markers of hepatitis viruses were evaluated in patients with different types of leukemia in comparing with control group. In this cross sectional study, 100 EDTA-treated blood samples were collected from leukemia patients and also from healthy control group, respectively. Serological and molecular markers of HBV, HCV and HDV viruses were analyzed for determination of the role of these hepatitis viruses in clinical outcomes of leukemia disorders. Increasing risk factors of leukemia were evaluated statistically in two studied groups by SPSS software. One of molecular and immunological markers of HBV, HDV and HCV was found in 24 of 100 (24%), 22 of 100 (22%), and 1 of 100 (1%) patients with leukemia and in 12 of 100 (12%), 6 of 100 (6%), and 2 of 100 (2%) control patients. Significant differences were detected in detection of HBsAg (P = 0.02), HBeAb (P = 0.009), and HCV-RNA (P = 0.05) between leukemia patients and control group, respectively. The high prevalence of HBV and HCV infective markers were detected in ALL and AML patients. Identification of high prevalence of HBV and HCV infective markers in leukemia patients proposed strong association between hepatitis viral infections and leukemia. Therefore, evaluation of the prevalence of viral hepatitis infections in larger groups of patients with long lasting follow up is suggesting.     

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.     

Seroepidemiology of hepatitis C virus infection in Japan and HCV infection in haemodialysis patients

Abstract: Since January 1990, Japanese Red Cross Blood Centres have introduced hepatitis C virus screening with a first-generation ELISA. From April to December 1992, approximately 0.98% among 10905 489 blood donations screened by a second-generation assay were anti-HCV-positive in all Japan. Seropositivity of anti-HCV increased with the age and serum transminase value in both sexes. In blood donors having a history of transfusion, the anti-HCV reactive rate was 7.4%. The results of the study made by the Japanese Red Cross Non-A, Non-B Hepatitis Research Group show the effectiveness of implementation of HCV screening to prevent posttransfusion hepatitis. Consecutive haemodialysis patients with chronic renal failure are at risk for inflection by a variety of blood-borne agents transmitted within dialysis units. Because of their immunocompromised state, they frequently also have an unusual susceptibility to a variety of nosocomial infections, such as HBV, and HTLV-I. We tested the prevalence of anti-HCV in 1423 (848 males and 575 females) haemodialiysis patients from 18 hospitals in Kumamoto Prefecture, Japan using the Orhto first generation anti- HCV screening assay. There were 316 patients (22.2%) positive for HCV antibodies. The second-generation test was positive in most haemodialysis patients who were eractive to the firs-generation assay. The prevalence of HCV infection increased with the duration of haemodialysis, yet there was a high frequency of HCV seropositivity even wihtout blood transfusion. Acquisition of HCV in dialysis patients could be explained by HCV seropositivity even without blood (all haemodialysis are done with disposable kits, and needles), by secondary HCV infection after the immunodeficiency of haemodialysis, or by HCV infection of the kidney or glomerular deposition of immune HCV/anti-HCV complexes leading to chronic renal failure (as with HBV infection of the liver and kidney).     

Transfusion-transmitted infectious diseases

A spectrum of blood-borne infectious agents is transmitted through transfusion of infected blood donated by apparently healthy and asymptomatic blood donors. The diversity of infectious agents includes hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency viruses (HIV-1/2), human T-cell lymphotropic viruses (HTLV-I/II), Cytomegalovirus (CMV), Parvovirus B19, West Nile Virus (WNV), Dengue virus, trypanosomiasis, malaria, and variant CJD. Several strategies are implemented to reduce the risk of transmitting these infectious agents by donor exclusion for clinical history of risk factors, screening for the serological markers of infections, and nucleic acid testing (NAT) by viral gene amplification for direct and sensitive detection of the known infectious agents. Consequently, transfusions are safer now than ever before and we have learnt how to mitigate risks of emerging infectious diseases such as West Nile, Chikungunya, and Dengue viruses.     

Hepatitis C prevalence and risk factors in hemodialysis patients in Central Brazil: a survey by polymerase chain reaction and serological methods

An hemodialysis population in Central Brazil was screened by polymerase chain reaction (PCR) and serological methods to assess the prevalence of hepatitis C virus (HCV) infection and to investigate associated risk factors. All hemodialysis patients (n=428) were interviewed in eight dialysis units in Goiania city. Blood samples were collected and serum samples screened for anti-HCV antibodies by an enzyme-linked immunosorbent assay (ELISA). Positive samples were retested for confirmation with a line immunoassay (LIA). All samples were also tested for HCV RNA by the PCR. An overall prevalence of 46.7% (CI 95%: 42-51.5) was found, ranging from 20.7% (CI 95%: 8.8-38.1) to 90.4% (CI 95%: 79.9-96.4) depending on the dialysis unit. Of the 428 patients, 185 were found to be seropositive by ELISA, and 167 were confirmed positive by LIA, resulting in an anti-HCV prevalence of 39%. A total of 131 patients were HCV RNA-positive. HCV viremia was present in 63.5% of the anti-HCV-positive patients and in 10.3% of the anti-HCV-negative patients. Univariate analysis of risk factors showed that the number of previous blood transfusions, transfusion of blood before mandatory screening for anti-HCV, length of time on hemodialysis, and treatment in multiple units were associated with HCV positivity. However, multivariate analysis revealed that blood transfusion before screening for anti-HCV and length of time on hemodialysis were significantly associated with HCV infection in this population. These data suggest that nosocomial transmission may play a role in the spread of HCV in the dialysis units studied. In addition to anti-HCV screening, HCV RNA detection is necessary for the diagnosis of HCV infection in hemodialysis patients.     

Clinical and epidemiologic characteristics of hepatitis C in a gastroenterology/hepatology practice in Ottawa.

OBJECTIVE: To examine the clinical and epidemiologic features of hepatitis C virus (HCV) infection in a gastroenterology/hepatology practice in Ottawa. DESIGN: Retrospective chart review. PATIENTS: Sixty-three consecutive patients found to be anti-HCV positive. Their charts were analysed with respect to risk factors, history of hepatitis, serum aspartate aminotransferase (AST) levels and the presence of hepatitis B markers. The long-term sexual partners of 29 patients agreed to undergo HCV antibody testing. RESULTS: Of the patients 48 (76%) had been exposed to HCV parenterally: 27 used intravenous drugs, and 21 had received blood or blood products. Eleven patients did not have any known risk factor (sporadic infection), but eight of them had lived in countries where hepatitis C may be more prevalent; the other three had locally acquired infection. The mean serum AST level at the first visit was 140 (normally less than 40) IU/L. At least one hepatitis B marker was identified in 33% of the patients. None of the sexual partners who were tested were anti-HCV positive. CONCLUSION: Most cases of hepatitis C in Ottawa are acquired through parenteral exposure; sexual transmission is rare. Sporadic infection in the Ottawa region is rare but may be more common in people from countries with a higher prevalence rate of hepatitis C. Most cases of hepatitis C are asymptomatic.     

Seroprevalence of hepatitis B and C virus markers among blood donors in Rio de Janeiro, Brazil, 1998-2005

The prevalence of infection by hepatitis B (HBV) and C (HCV) viruses varies among geographical regions. In order to determine the prevalence of HBV and HCV infection in voluntary blood donors we evaluated the prevalence of HBsAg, anti-HBc, and anti-HCV markers of 128,497 blood donor samples collected from 1998 to 2005 in the state of Rio de Janeiro. These markers were analyzed by immunoenzymatic tests, as determined by the Ministry of Health. Data were obtained from the Sorology Laboratory of the Hemotherapy Service of the Instituto Nacional de Cancer, Rio de Janeiro. Overall prevalence estimates were: 0.27% for HBsAg, 3.68% for anti-HBc, and 0.90% for anti-HCV. There was a significant decrease in the overall prevalence of HBsAg (from 0.36 to 0.14%) and anti-HBc (from 6.12 to 2.05%) in the period encompassed between 1998-2005. Similarly, there was a decline in anti-HCV prevalence rates in Brazilian blood donors, from 1.04% in 1998 to 0.79% in 2004, with an increase of HCV prevalence to 1.09% in 2005. These prevalence estimates were higher than those found in other countries, indicating high rates of infection by HBV and HCV and a persistent risk of HBV and HCV transmission by transfusion.     

The occurrence rate of HGV/GBV-C RNA and risk factors in patients of narcological dispensary in Novosibirsk

The occurrence rate of HGV/GBV-C RNA, genotypic variety of isolates and various risk factors of infection with HGV/GBV-C were evaluated in 500 patients of the narcological dispensary of Novosibirsk. The occurrence rate of HGV/GBV-C RNA among all examined blood sera was 33.6%. At the same time in blood sera with HCV markers the occurrence rate of HGV/ GBV-C was 42.9% and in sera with negative results for markers HCV--25%. For gene typing of obtained isolates the direct sequencing of the amplification products of fragment NS3B and the phylogenetic analysis of the sequences thus obtained were used. Almost all isolates subjected to gene typing belonged to genotype 2, widespread in Europe, and only 1 isolate was classified with genotype 4. Statistically significant (p<0.05) risk of HGV/GBV-C infection among the examined subjects was linked with the intravenous use of drugs (OR 2.15), risky sexual behavior (OR 1.8) and the presence of virus hepatitis C (OR 2.26).     

Occurrence of markers, distribution of genotypes and risk factors for viral hepatitis C among some groups of the population in the Novosibirsk region

The occurrence of markers, genotypic variability of isolates and risk factors for viral hepatitis C (HCV) were studied in 4 groups of residents of the Novosibirsk region (altogether 2,000 persons). Anti-HCV IgG were detected within the range from 4.6% among medical personnel to 48% among the patients of the drug-abuse clinic. The detection rate of HCV RNA in seropositive samples varied from 79.3% to 86.3%. The determination of genotype was carried out for 388 isolates: 1b--50.3%, 2a--4.4%, 2c--0.3%, 3a--44.8%. The highest risk indices with respect to HCV among the residents of the region were linked with the drug use (OR=77.5; p<0.05) as well as with risky behavior and low social status. The elevated numbers of seropositive persons were detected among unemployed (OR=16.3), alcohol abusers (OR=3.9), persons having more than 4 sex partners in their lifetime (OR=4.3) and persons having homosexual contacts (OR=6.6). In some groups blood transfusions also played a definite role in the transmission of HCV. In the analysis, carried out separately for two different genotypes the intravenous use of drugs was perceptibly stronger linked with VHC of genotype 3 (OR=85.5) in comparison with HCV of genotype 1 (OR=49.3) and genotype 2 (OR=41.1). Genotype 1 prevailed in the older age group and genotype 3, among young people.     

Prevalence, genotypes and risk factors associated with hepatitis C virus infection in hemodialysis patients in Campo Grande, MS, Brazil

A survey was conducted among the hemodialysis units of the city of Campo Grande, located in the state of Mato Grosso do Sul in the Mid-west region of Brazil, with the aim of investigating the prevalence, risk factors, and genotypes of hepatitis C virus (HCV) infection. A total of 163 patients were interviewed in five dialysis units. Serum samples were screened for anti-HCV. Positive samples were tested for HCV RNA and genotyped. The prevalence of anti-HCV was 11% (95% CI: 6.8-17.1). A history of transfusion with blood that was not screened for anti-HCV and length of time on hemodialysis were associated with HCV infection. HCV RNA was detected in 12 samples: ten were of genotype 1, subtypes 1a (75%) and 1b (8.3%), and two were of genotype 3, subtype 3a (16.7%).     

Investigation of putative multisubtype hepatitis C virus infections in vivo by heteroduplex mobility analysis of core/envelope subgenomes

The frequency that multiple different subtypes of hepatitis C virus (HCV) simultaneously infect a given individual is controversial. To address this question, heteroduplex mobility analysis (HMA) of portions of the HCV core and envelope 1 region was optimized for sensitive and specific detection of mixtures of HCV genomes of different genotype or subtype. Using the standard HCV genotyping approach of 5'-untranslated region (UTR) analysis, 28 of 374 (7.5%) chronic hepatitis C research subjects were classified as having either multiple-subtype HCV infections (n = 21) or switching HCV subtypes over time (n = 7), the latter pattern implying viral superinfection. Upon retesting of specimens by HMA, 25 of 28 multiple-subtype results could not be reproduced. All three patients with positive results were injection drug users with potential multiple HCV exposures. To address the hypothesis of tissue sequestration of multiple-subtype HCV infections, liver (n = 22), peripheral blood mononuclear cell (n = 13), perihepatic lymph node (n = 16), and serum (n = 19) specimens from 23 subjects with end-stage hepatitis C were collected and analyzed by the HMA technique. Whereas 5'-UTR results implicated mixed-subtype HCV infections in 2 subjects, HMA testing revealed no evidence of a second HCV subtype in any tissue compartment (0 of 70 compartments [0%]) or within any given subject (0 of 23 subjects [0%]). In summary, a large proportion of mixed-genotype and switching-genotype patterns generated by 5'-UTR analysis were not reproducible using the HMA approach, emphasizing the need for additional study.     

A cDNA fragment of hepatitis C virus isolated from an implicated donor of post-transfusion non-A, non-B hepatitis in Japan.

Recently, a cDNA from the hepatitis C virus (HCV) RNA genome has been isolated in the USA from a chronically infected chimpanzee. In order to isolate HCV cDNA derived from human material, RNA was extracted from plasma of a Japanese blood donor implicated in post-transfusion non-A, non-B hepatitis and HCV cDNA was synthesized and amplified by the PCR method using HCV-specific oligonucleotide primers. The cDNA fragment, 583 nucleotides long, showed 79.8% homology at the nucleotide level and 92.2% homology at the amino acid level compared with the prototype HCV cDNA. These results provides further evidence to show that HCV is closely associated with the development of post transfusion non-A, non-B hepatitis.     

Hepatitis C virus prevalence among an immigrant community to the southern Amazon, Brazil

A community-based random survey was conducted in a southern Brazilian Amazonian county aiming to investigate hepatitis C virus (HCV) infection prevalence and the association of demographic variables and lifestyle behaviours. Seven hundred eighty individuals were serologically screened with a third generation enzyme-linked immunosorbent assay to detect anti-HCV antibodies between 1994/1995. Positive samples were retested for confirmation with a line immunoassay (LIA, Inno-LIA HCV Ab III). Most of these subjects were low income and came from southern Brazilian states (65.8). Two point four percent (IC 95% 1.2%- 4.6%) of the subjects had LIA-confirmed anti-HCV antibodies reactivity. The age-specific prevalence of HCV antibodies slightly increased with age, with the highest prevalence after the age of 40 years. The results of multivariate analysis indicate a strong association between HCV antibodies and previous surgery and history of intravenous drug use. There were no apparent association with gender, hepatitis B virus markers, blood transfusion, and sexual activity. Mean time living in Amazon did not differ between confirmed and negative anti-HCV individuals. The present data point out an intermediate endemicity of HCV infection among this immigrant community to the Amazon region and that few HCV infected participants presented known risk factors.     

Tendency and analysis of the epidemic situation in parenteral virus hepatitis B and C in the Russian Federation and a number of its regions

As revealed in the present survey, during the last 3 years, against a background of decreased number of registered cases of acute hepatitis B (HB) and acute hepatitis C (HC), an increase in the proportion of patients with the chronic forms of these diseases was observed. The incidence rate of carriership of hepatitis B (HBV) and hepatitis C viruses (HCV) is many times greater than morbidity rates in acute and chronic forms of the disease. Such differences could be due to imperfect laboratory and clinical diagnosis. The registered statistics on HBV and HCV carriership included newly detected HBsAg and anti-HCV in the absence of clinical manifestations, which did not reflect the true spread of HBV and HCV in a given territory. The group of HBV and HCV carriers was found to include a considerable proportion of patients with asymptomatic form of HB and HC. It was testing for HBsAg, anti-HCV only without determination of virus replication markers (anti-HBc IgM, HBV DNA, anti-HCV IgM, HCV RNA) that seemingly determined the category of carriers greatly exceeding the true incidence. To obtain reliable epidemiological information, the complex detection of HB and HC infection markers is necessary.     

Decline of hepatitis C infection in hemodialysis patients in Central Brazil: a ten years of surveillance

Hepatitis C virus (HCV) has been a significant problem for hemodialysis patients. However this infection has declined in regions where the screening for anti-HCV in blood banks and hemodialysis-specific infection control measures were adopted. In Brazil, these measures were implemented in 1993 and 1996, respectively. In addition, all studied units have implemented isolation of anti-HCV positive patients since 2000. In order to evaluate the impact of these policies in the HCV infection prevalence, accumulated incidence, and risk factors in hemodialysis population of Goiania City, Central Brazil, all patients were interviewed and serum samples tested for HCV antibodies in 1993, 1996, 1999, and 2002. In the first six years (1993-1999), anti-HCV prevalence increased from 28.2 to 37.2%, however a b decrease in positivity was detected between 1999 and 2002 (37.8 vs 16.5%) when the measures were fully implemented. Also, a decrease of the anti-HCV accumulated incidence in cohorts of susceptible individuals during 1993-2002 (71%), 1996-2002 (34.2%), and 1999-2002 (11.7%) was found. Analysis of risk factors showed that length of time on hemodialysis, blood transfusion before screening for anti-HCV and treatment in multiple units were statistically associated with anti-HCV (p < 0.05). Our study showed a significant decline of hepatitis C infection in hemodialysis patients of Central Brazil, gratifying the importance of public health strategies for control and prevention of hepatitis C in the hemodialysis units.     

Cellular immune responses associated with occult hepatitis C virus infection of the liver

Occult hepatitis C virus (HCV) infection is a type of recently identified chronic infection that is evidenced only by detection of HCV RNA in liver; patients consistently test negative for antibodies to HCV and HCV RNA in serum. Using ex vivo and in vitro measures of T-cell responses, we have identified functional virus-specific memory CD4(+) and CD8(+) T cells in the peripheral blood of patients with occult HCV infection. The features of the virus-specific T cells were consistent with immune surveillance functions, supporting previous exposure to HCV. In addition, the magnitudes of CD4(+) and CD8(+) T-cell responses were in parallel and correlated inversely with the extent of liver HCV infection. The detection of HCV-specific T cells in individuals in whom HCV RNA can persist in the liver despite the absence of viremia and antibodies indicates that HCV replication is prolonged in the face of virus-specific CD4(+) and CD8(+) T-cell responses. These findings demonstrate that HCV-specific cellular immune responses are markers not only of previous exposure to and recovery from HCV but also of ongoing occult HCV infection.     

Hepatitis C virus-RNA, immunoglobulin M anti-HCV and risk factors in haemodialysis patients

In order to evaluate hepatitis C virus-RNA (HCV-RNA), immunoglobulin M (IgM) anti-HCV and risk factors in haemodialysis patients, 180 subjects (45 HCV negative and 135 HCV positive) were studied. Sex, age, duration of dialysis, number of transfusions and ALT were also considered. HCV-RNA was determined by the Amplicor HCV test, and IgM anti-HCV by the Abbott HCV IgM EIA. These markers were present in 40% and 30.4% of anti-HCV positive subjects. The agreement between the two tests employed was 77%. The results showed a close association between HCV-RNA and IgM anti-HCV with abnormal ALT levels and between HCV-RNA and the number of transfusions. Both of these markers were different when correlated with age and time on dialysis, respectively. Therefore, IgM anti-HCV may also serve as a serological marker of HCV infection and a complementary marker of virus replication.     

The first large-scale nucleic acid amplification testing (NAT) of donated blood using multiplex reagent for simultaneous detection of HBV, HCV, and HIV-1 and significance of NAT for HBV.

The first nationwide nucleic acid amplification testing (NAT) for hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type 1 (HIV-1) of voluntarily donated blood after serological pre-screening and before release of cellular components and plasma for fractionation was implemented by the Japanese Red Cross Blood Transfusion Services. From February 1, 2000 to April 30, 2001, specimens from 6,805,010 units of serologically negative donation were screened in minipools of 50 samples within 24 hr after blood donation by NAT using multiplex HBV/HCV/HIV-1 reagent for blood transfusion including short shelf-life platelets. Among them, 112 HBV DNA-positives, 25 HCV RNA positives and 4 HIV-1 RNA positives were screened out and we could prevent transfusion of these NAT positive units. Subtypes/genotypes of HBV DNA, adr/C, adw/A, adw/B, adw/C, ayr/C and ayw/D were found and adr/C was predominant. A total of 61.6 % of them (69/112) were negative by overnight EIA. Sixth three of HBV NAT-positive samples carried virus loads less than 10(4) copies/mL and 92.1 % of them (58/63) were negative by overnight EIA. The virus growth curves of HBV in 6 cases obtained by retrospective and prospective follow-up study showed exponential straight lines in the early stage of serological window periods and the log times of HBV growth (10 fold increase) in serological window period were between 4.6 and 7.6 days. NAT screening with highly sensitive reagents in pool of specimens is useful to exclude blood units with low level of HBV and HBV mutants from blood transfusion.     

Prevalence of human immunodeficiency virus infection in alcoholics

To verify the prevalence of infection by human immunodeficiency virus (HIV) in alcoholics we studied 131 alcoholic patients (119 males and 12 females) with a mean age of 44.3 +/- 10.8 years. Serum samples were collected from this group and analysed, by ELISA, for antibodies against HIV as well as for serological markers for hepatitis B virus (HBV) and hepatitis C virus (HCV). As we have previously described, we found a high prevalence of HBV (26.4%) and HCV (4.2%) markers as compared to the prevalence of these markers in samples of normal blood donors from Uberlandia's Hemocentro Regional, which are 4% and 0.4%, respectively. Of the 131 patients, four (3%) had antibodies against HIV, three (75%) of which were injecting drug users (IDU). In the HIV-negative group, only one patient was an IDU. The prevalence of HIV in our population, according to data from the city's Health Secretary, varies from 3.1% to 6.2%. We conclude that, at least for the moment, alcoholism per se, did not constitute an important risk factor for HIV infection. However, acquired immunodeficiency syndrome is a rather recent disease as compared to hepatitis B and C and, as the transmission routes are similar for HIV and hepatitis viruses, an increase in the incidence of HIV infection in alcoholics may be just a question of time.