Cases Of Post-transfusion Hepatitis Research Articles

Molecular Detection of Hepatitis B Virus Among HBsAg Non-Reactive Blood of Donors

Despite all blood donations being tested routinely for HBsAg as a clinical marker of transmissible HBV, cases of post-transfusion hepatitis B virus infection are still being reported because molecular studies using Polymerase Chain Reaction (PCR) are not routinely available for Transfusion Transmissible Infection (TTIs) testing. In this study, we sought to use the PCR technique to re-screen donated blood that had already been proven to be HBsAg non-reactive with rapid diagnostic testing and ELISA. One hundred and eighty-five samples were obtained from a proportion of the blood deposited at the blood bank of the Federal Medical Center, Birnin-Kebbi, Nigeria. Socio-demographic parameters such as age group, status, ethnicity, occupation, and group PCV were obtained from donors' records. Nested PCR was employed to detect HBV DNA. Furthermore, genotyping was performed to determine HBV genotypes in the positive samples using PCR with genotype-specific primers. Of the 185 donors, it was observed that five (2.7%) of the population were positive for HBV. HBV is more common among people aged 18–30, singles, Hausa, and self-employed. In addition, the five positive samples were of genotype E. This study suggests the need to complement antibody-based tests with DNA testing for effective HBV screening and consequent safe transfusion. Keywords: Hepatitis B Virus; HBsAg; PCR; DNA testing

Screening of Occult Hepatitis B Virus Infection among Egyptian Blood Donors.

Background:Although sensitive screening assays for hepatitis B virus surface antigen (HBsAg) are available, special cases of post-transfusion hepatitis B virus infection still occur. The present study was conducted to evaluate the prevalence of anti-hepatitis B core (anti-HBc) positivity and the presence of HBV-DNA in serum samples of healthy blood donors negative for both HBsAg and anti-HCV antibodies in Benha, Egypt. Materials and methods: The study included a screening of 450 selected blood donors. The distribution of blood donors was 288 males (64%) and 162 females (36%). The recruited blood donors who met the criteria for blood donation were routinely screened for HBsAg, HIV I/2-Ab, and syphilis-antibodies. The blood units for donations were further analyzed for the presence of HBc-IgM and HBV-DNA levels by PCR method. Results: Testing of the accepted units for the donation was about 12 (2.7%) HBc-IgM positive, and 9 (2%) HBV-DNA positive units. The standard screening of blood unit failed to recognize early acute or window HBV infection where HBsAg is missing. Conclusions:Our investigations proposed that sensitive methods for the detection of HBV by PCR might be recommended in the screening of donated blood. Furthermore, anti-HBc antibodies should be tested regularly on all blood donation units.

Socio-demographic and other risk factors associated with HBV potential infectivity among Hepatitis B Surface Antigen Negative blood donors

Background: There have been cases of post-transfusion hepatitis in spite of transfusion of hepatitis B surface antigen negative blood unit. Objective: To determine the socio-demographic and risk factors associated with positivity of anti-HBc among Hepatitis B surface antigen negative blood donors. Methods: A cross-sectional study was carried out among 490 HBsAg negative blood donors. The participants were tested for anti-HBc using ELISA kits. A structured questionnaire was used to obtain socio-demographic data and other possible risk factors. Results: The mean age of the participants was 32.5 ± 9.5 years with male predominance of 462 (94.3%) while 375 (76.5%) were first time donors. Of the 490 HBsAg negative blood donors, 89 (18.2%) were positive for anti-HBc. The highest prevalence of anti-HBc occurred in the 26-34 years age group. Independent risk factors associated with the presence of anti-HBc included age (p = 0.049), marital status (p = 0.003), dental and surgical procedure (p<0.001), traditional practices such as tattoo, body piercing and scarification (p<0.001) and previous blood transfusion (p = 0.016). Conclusion: Age, marital status, traditional and cultural practices like scarifications, tattoo and body piercing and history of blood transfusion were risk factors associated with HBV infection despite HBsAg negativity.

Hepatitis C: Viral Features and Clinical Aspects

Knowledge on hepatitis caused by hepatitis C virus (HCV) has increased since 1989, when the virus was identified and cloned from the complementary deoxyribonucleic acid (DNA) extracted from cells of an experimentally infected chimpanzee with the virus of hepatitis non-A, non-B. Approximately 90% of cases of post-transfusion hepatitis and 50% to 70% of sporadic cases of hepatitis non-A and non-B were caused by HCV. Thus, the discovery of this virus has enabled the development of serological diagnosis techniques. In recent years, there have been major advances in the characterization of the molecular structure of HCV, the development of diagnostic tests with high specificity and sensitivity, knowledge about the pathogenic mechanisms and improved therapeutic options. The continued study of this disease allowed the characterization of its epidemiology and main routes of transmission. Currently, HCV competes with alcoholic liver disease as the leading cause of chronic liver disease. As shown asymptomatic in most cases, this disease progresses silently for chronicity with a consequent increase in cases of liver cirrhosis and hepatocellular carcinoma (HCC). This study reviews the main aspects of the hepatitis C. HCV infection is progressive, silent and causes the patient serious complications such as cirrhosis and HCC, and brings high costs to the government. Therefore, given the diversity of problems caused by epidemic HCV, this study is essential to new discoveries about this health problem.

Blood component donors and recipients as linked study objects in epidemiological population-based surveys

A population of hematological cancer patients as recipients of many blood components and that of donors of blood components and bone marrow are related to the common event of contamination with viruses of blood-borne infections; which occurs and is detectable during long-term treatment and follow-up. They share interaction traits and diverse communication mechanisms, which call for complex interrelated trials in both groups with a mandatory epidemiological evidenced-based investigation of all cases of posttransfusion hepatitis B and/or C. The identity of infection with hepatitis B and C viruses, human immunodeficiency virus, and their association should be simultaneously studied in the populations of both donors and recipients of blood components and bone marrow.

A Neutralization Epitope in the Hepatitis C Virus E2 Glycoprotein Interacts with Host Entry Factor CD81

The identification of a specific immunogenic candidate that will effectively activate the appropriate pathway for neutralizing antibody production is fundamental for vaccine design. By using a monoclonal antibody (1H8) that neutralizes HCV in vitro, we have demonstrated here that 1H8 recognized an epitope mapped between residues A524 and W529 of the E2 protein. We also found that the epitope residues A524, P525, Y527 and W529 were crucial for antibody binding, while the residues T526, Y527 and W529 within the same epitope engaged in the interaction with the host entry factor CD81. Furthermore, we detected “1H8-like” antibodies, defined as those with amino acid-specificity similar to 1H8, in the plasma of patients with chronic HCV infection. The time course study of plasma samples from Patient H, a well-characterized case of post-transfusion hepatitis C, showed that “1H8-like” antibodies could be detected in a sample collected almost two years after the initial infection, thus confirming the immunogenicity of this epitope in vivo. The characterization of this neutralization epitope with a function in host entry factor CD81 interaction should enhance our understanding of antibody-mediated neutralization of HCV infections.

Occult HBV感染と考えられる血液製剤による輸血後B型急性肝炎の一例

輸血によるB型急性肝炎の一例を報告する．症例は50歳代男性．心臓手術を受けた際に多量の輸血を受けた．輸血約3カ月後にはHBs抗原，HBs抗体，HBc抗体いずれも陰性であったが，6カ月後に肝障害を認めた．HBs抗原陽転化および血中HBV-DNA陽性によりB型急性肝炎と診断された．安静のみで肝炎は改善し，血中HBV-DNAは数週間で検出不能となった．輸血製剤の調査により新鮮凍結血漿の1本でHBV-DNAが検出され，塩基配列が患者から得られたウイルスのものと一致した．献血者は頻回に献血を行っておりOccult HBV感染と考えられた．核酸増幅検査の精度向上にかかわらず，Occult HBV感染者由来と思われる輸血後B型肝炎は年間数例の報告が続いている．2012年8月にHBc抗体低力価かつHBs抗体低力価の献血血液を不使用とする新たな対策が講じられ，感染者の減少が期待されている．

Hepatitis B virus traceback and lookback: factors to consider

Hepatitis B virus traceback and lookback: factors to consider

Transfusion‐transmitted hepatitis E in a ‘nonhyperendemic’ country

Indigenous hepatitis E is increasingly recognized in developed countries, where it may be a zoonosis. We describe the first case of transfusion-transmitted hepatitis E in the UK from a blood donor who had no history of recent travel abroad. Follow-up of the donor and recipients of the blood products was carried out using serological and molecular techniques. Acute hepatitis E was transmitted to one of two recipients. The infected patient would have received a larger volume of the donor's plasma. HEV subgenomic sequences carried by the donor and recipient were identical. This is the first case of post-transfusion hepatitis E in the UK. Secondary transmission of hepatitis E indigenous to a nonhyperendemic country may occur by blood transfusion. It is important that blood donors inform the transfusion service of all post-donation illnesses so that appropriate interventions can take place.

Replacement therapy with virus-inactivated clotting factor concentrates in patients with severe hemophilia in Heidelberg.

As early as 1943, Beeson had observed a connection between the transfusion of blood products and the emergence of hepatitis.[1] Jaundice developed in 7 patients 1 to 4 months after the administration of whole blood or plasma. Further reports on the development of hepatitis in hemophilia patients were published in the mid-1970s.[2] [3] [4] Today, it is generally accepted that the hepatitis C virus (HCV) causes most cases of posttransfusion hepatitis in the western world. It is responsible for most of the chronic liver disorders observed in hemophilia patients treated with clotting factor concentrates. The HCV infection can be associated with replacement therapy involving clotting factor concentrates administered in the early 1970s.[2] [5] [6]

Hepatitis C lookback

Hepatitis C was responsible for the majority of cases of posttransfusion hepatitis before the introduction of a specific screening test for blood donors. Infected recipients may remain asymptomatic for many years, but cirrhosis and hepatocellular carcinoma may develop decades after infection. Lookback, or the identification of recipients of potentially contaminated blood, is now being conducted in many countries, including Holland, Denmark, the United Kingdom, Canada, New Zealand, and the United States. In targeted lookback, recipients of blood from donors subsequently found to be positive for hepatitis C are notified and advised to undergo testing. In general lookback, all patients who received blood before being tested for hepatitis C are advised to undergo testing. Difficulties with both forms of lookback illustrate the importance of vein-to-vein tracking of blood products, including the potential utility of a centralized registry of blood product recipients.

Detection of hepatitis B virus DNA in blood units with anti-HBc as the only positive serological marker.

Serum samples from 10,629 blood donors were screened for hepatitis B virus (HBV) serological markers (HBsAg, anti-HBs, anti-HBc, anti-HBc IgM), anti-HCV, anti-HIV1/2 and ALT. Seventy five (0.7%) blood donors were found HBsAg-positive, 1,543 (14.5%) were carrying both anti-HBc and anti-HBs. whereas 507 (4.8%) samples were positive only for anti-HBc. Among the group of 507 anti-HBc positive samples, 303 were obtained from regular volunteer blood donors who were studied in two separate time intervals of at least 6 months' duration, and 204 were from first-time blood donors. The possibility of post-transfusion hepatitis B after donation of these 507 blood units was studied by determining the presence of HBV DNA as a marker of viral replication and infectivity. HBV DNA was detected by two methods (i) a chemiluminescent molecular hybridization assay, (ii) polymerase chain reaction (PCR) followed by DNA enzyme immunoassay (DEIA). Six out of 507 samples exhibited HBV DNA results in the gray zone of the hybridization assay, but were confirmed as negative by PCR DEIA. The other 501 samples were HBV DNA-negative by both methods, although 36 of them had increased ALT levels. No cases of post-transfusion hepatitis B were reported during the year in which these 501 blood units were provided. These results show that blood units which were positive only for anti-HBc, with normal ALT and were HBV DNA-negative may be considered not infectious for hepatitis B. Gray zone results of HBV DNA using hybridization quantitative assay must be confirmed as positive or negative by a more sensitive method such as PCR. Blood units which are anti-HBc-positive, with increased ALT levels and are HBV DNA-negative, which appear to not be related to HBV replication and infectivity, may be not safe for donation because of the potential existence of other as yet unknown, hepatotropic viruses.

Molecular analysis of HCV type 1 to 5 envelope gene: application to investigations of posttransfusion transmission of HCV.

Until 1990, HCV infection was common in transfused patients, resulting in more than 200,000 cases of posttransfusion hepatitis C in France alone. A molecular method that permits the investigation of posttransfusion hepatitis C infections is presented. Viral sequences in the envelope region of HCV were obtained for 12 pairs of blood recipients and their respective blood donors. The HCV strains studied belonged to types 1 (subtypes 1a and 1b), 2, 3, 4, and 5. Genetic distances and mutation rates were determined, and sequences were submitted to phylogenetic analysis along with sequences retrieved from nucleotide databases. Pairwise distances in the donor-recipient pairs were found to be less than 0.05 mutation per site, which corresponds to a mutation rate ranging from 0.6 x 10(-3) to 2.1 x 10(-3) per site per year. Sequences obtained from the 12 donor-recipient pairs clustered in 12 monophyletic nests. The genetic analysis of the envelope region of HCV can be used for the forensic evaluation of virus transmission. It permits the refutation of a link between blood transfusion and HCV transmission, rather than proof of the existence of such a link.

Optimal blood donation screening annotation.

Optimal blood donation screening annotation.

TTV - a virus searching for a disease.

TTV is a new virus which was identified in the serum of a patient with non-A-G post-transfusion hepatitis in Japan. The original workers aimed to account for the small number of post transfusion hepatitis cases found in their clinical practice. Subsequent work has attempted to determine the properties and unravel the natural history of the new agent. The original study applied representational difference analysis to detect foreign DNA sequences which were present only during the acute phase of illness. Subsequent studies have used PCR to study the agent in serum liver and faeces. This review summarises the published data from clinical and epidemiological studies in different countries. The inclusion of the virus in the parvovirus family seems premature because its size is unknown, its reported density is too light and its sequence lacks the characteristic long terminal repeats. The agent can be found in 1-40% of health blood donors in different countries and also in faeces. TTV is ubiquitous but its taxonomic identity and disease load remain to be determined.

Hepatitis C virus infection: immune responsiveness and interferon-alpha treatment.

Hepatitis C virus (HCV) is responsible for most cases of posttransfusion hepatitis and sporadic or community-acquired non-A, non-B hepatitis. Different generations of enzyme-linked immunosorbent assay have been generated for detecting antibodies to HCV epitopes. HCV-RNA quantitative analysis has been developed by means of polymerase chain reaction technique. This approach is the only reliable method for HCV-RNA tissue localization, being helpful in early diagnosis. HCV infected liver is characterized by an inflammatory infiltrate including CD4+, CD8+, and B lymphocytes. Evidence has been provided that in HCV patients CD8+ cell response is associated with low level of viraemia and higher level of disease activity. CD4+ T cells exhibit specificity for the core antigen, also correlating with disease activity and viraemia. Costimulatory molecules, cytokines, oxygen radicals, the complex Fas/Fas-ligand and soluble class I HLA structures are discussed as putative cofactors involved in disease evolution. Various forms of interferon (IFN)-alpha have been evaluated for the treatment of patients with HCV infection. Initial enthusiasm has been attenuated by the evidence of a low sustained virological response rate and the constant side effects of IFN-alpha therapy in patients with chronic HCV disease. Among possible markers for predicting therapeutic outcome in HCV-positive individuals, anti-core antibodies correlate positively with response to IFN-alpha administration, as well as reduction of interleukin-2 serum levels has been detected in patients with a good therapeutic response. Association between HCV infection and autoimmune phenomena, also in relation to IFN-alpha therapy has been reported. Finally, results of the combined treatment with IFN-alpha/ribavirin are illustrated.

Characterization of the humoral and cellular immune responses against hepatitis C virus core induced by DNA-based immunization

Hepatitis C Virus (HCV) causes most cases of posttransfusion hepatitis. Chronic HCV infection is highly related to chronic hepatitis, cirrhosis and hepatocellular carcinoma. Current therapies are only minimally effective and no vaccine has been developed. DNA-based immunization could be of prophylactic and therapeutic value for HCV infection. By intramuscular inoculation in BALB/c mice with an HCV recombinant plasmid pCI-HCV-C, we found significant levels of IgM antibody, but no significant IgG rise. After boost the immunized mice with recombinant HCV-core protein (cp1–10; 1–164aa), the anticore IgG, verified by Western-blotting, rose rapidly, which was two weeks earlier than that with control plasmid. Spleen cells from pCI-HCV-C immunized mice gave higher proliferation index (PI) than control ( P<0.05). The PI of cp1–10 boosted mice was even higher. Proliferation blocking assay with mAb proved the responding cell to be of CD4 +CD8 − phenotype, supporting specific priming of T helper cells. A 51Cr-releasing CTL assay specific for HCV-core was developed, and a specific CTL response against HCV-core was demonstrated in both pCI-HCV-C immunized mice and mice boosted with cp1–10. Strong cytotoxic activity against peptide-pulsed p815 cells (H-2 d), but not EL-4 cells (H-2 b), suggested MHC class I restriction of the CTL activity. Blocking of CTL with mAb proved the effector cells to be of CD4 −CD8 +. Three CTL epitopes in HCV-core protein were demonstrated. We failed to detect CTL when immunized only with core protein. The results suggested that vaccination with HCV-core derived DNA sequences could be an effective method to induce humoral and cellular immune responses to HCV.

ＨＣＶ感染早期のＨＣＶ抗体陰性時期での献血に由来する輸血後Ｃ型肝炎の１例

ＨＣＶ感染早期のＨＣＶ抗体陰性時期での献血に由来する輸血後Ｃ型肝炎の１例

Detection of antibodies to hepatitis C virus in seronegative patients using an immune complex dissociation assay.

Sera from a small percentage of hepatitis C virus (HCV)-infected blood donors do not react in the currently available assays for detection of antibody to HCV (anti-HCV) and, as a consequence, hepatitis C may develop in recipients of this blood. One possible explanation for this phenomenon is that antibody is present but cannot be detected because it is sequestered in circulating immune complexes. To test this hypothesis, an immune complex dissociation (ICD) assay was developed to disrupt any immune complexes that might be present in these anti-HCV-negative, HCV RNA-positive sera. A positive result in this test would indicate that antibody is present in these patients but is not detectable under routine anti-HCV testing conditions. Nine chronic and two acute HCV patients, all negative for antibody but positive for HCV RNA by reverse transcriptase-polymerase chain reaction (RT-PCR) were tested, together with appropriate controls. Three of the nine study patients with chronic HCV had evidence of anti-HCV after immune complex dissociation compared with none of the two patients with acute HCV. Although the number of patients tested was small, the negative results in the patients with acute HCV presumably indicates that anti-HCV seroconversion had not yet occurred. Incorporation of an ICD step into existing anti-HCV assays may enable blood banks to detect those rare instances of patients with chronic HCV who are antibody negative; this would minimize potential cases of post-transfusion hepatitis in recipients.

The effects of two infusion rates on platelet count and corrected count increment in pediatric oncology patients

To clarify the role of hepatitis-A virus (H.A.V.) in the aetiology of post-transfusion hepatitis unrelated to hepatitis-B virus, we have tested and titred pre-transfusion and convalescent serum samples from 32 patients for antibody to hepatitis-A antigen (anti-HA) by quantitative immune adherence hæmagglutination. 12 patients had no detectable anti-HA in either pre-transfusion or late convalescent serum; the other 20 had anti-HA in pretransfusion serum and no significant change in titre during convalescence. This study excludes. H.A.V. as the agent responsible for these cases of post-transfusion hepatitis and supports the existence of "non-A, non-B" hepatitis virus(es).