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The transmission of viral and non-viral infectious pathogens continues to be the most serious of the potential adverse effects of allogenic tissue transplantations. EU Directive 2006/17/EC stipulates that cadaveric blood specimens for serology testing in the context of post-mortem tissue donation must be taken not later than 24 h post-mortem. An expanded time slot would significantly improve the availability of tissue donations, but there are no significant data on the stability of infectious serology assays for anti-human immunodeficiency virus (HIV), anti-hepatitis C virus (HCV), hepatitis B virus (HBV) surface antigen (HBsAg) and anti-HBC core antigen (HBc) in samples collected more than 24 h post-mortem. In this prospective study, serum samples of 30 deceased persons were taken upon admission to the Institute of Forensic Medicine (University Hospital Hamburg-Eppendorf, Germany) and at 12, 24, 36 and 48 h post-mortem. All samples were measured twice, first using the Abbott AxSYM system, and then after ~9 months of storage at −70 °C using the BEP III System with Siemens and Ortho reagents. For HIV, six deceased persons with a pre-mortem HIV history were included. All samples (at committal and at 12, 24, 36, 48 h) were reactive. Indeterminate or false-negative results did not occur. For HCV, 17 deceased persons with a pre-mortem HCV history were included; 16 samples were reactive up to 48 h and one was reactive at 36 h post-mortem (48 h sample was not available). Indeterminate or false-negative results did not occur. For HBV, nine deceased persons were included: five samples were initially positive for HBsAg and remained positive up to 48 h, and eight of the samples were reactive for anti-HBc up to 48 h and one up to 36 h post-mortem (48 h sample was not available). Indeterminate or false-negative results did not occur. These data suggest that infectious serological testing may be extended for blood samples of potential tissue donors collected up to 48 h post-mortem to detect antibodies or antigens for HIV, HBV and HCV.

To evaluate the risk of hepatitis C virus (HCV) transmission via breast milk, we collected 76 samples of breast milk from 73 chronically HCV-infected women and serum samples from their 76 perinatally HCV-exposed children. Enzyme immunoassay and strip immunoblot assay were used for detection of antibodies to HCV, and reverse transcriptase-polymerase chain reaction analysis was used for detection of HCV RNA. None of the 76 samples of breast milk contained HCV RNA, whereas 37 (59.7%) of 62 mothers tested for HCV RNA had HCV viremia. Only 1 of the 76 breast-fed infants had evidence of HCV infection. Because HCV infection in this child was detected 1 month after birth, it seems unlikely that it was transmitted by breast-feeding. These results indicate that HCV infection in pregnant women should not be a contra-indication for breast-feeding.

It is important to shorten the window period after acute HIV infection in which infected individuals are still antibody-negative, especially in blood donors. Newly developed fourth-generation assays detect antibodies to HIV-1, including subtype O, and to HIV-2 and, simultaneously, p24 antigen of HIV-1. To evaluate this assay for daily routine work we compared it with different third-generation assays using sera from uninfected patients and patients with known HIV infection. The most interesting sera are those drawn during seroconversion from freshly infected patients. Whenever we encounter such a patient with acute HIV infection we store the serum in aliquots at -20 degrees C. Thus, we were able to establish our own seroconversion panel and use it in our laboratory for evaluation of new assays. The new test was shown to be able to detect all chronically HIV-infected individuals and four of six patients during seroconversion although in two of these patients conventional assays for HIV antibodies were still negative. The rate of unspecific reactivities was slightly higher as compared with third-generation assays.

It is important to shorten the window period after acute HIV infection in which infected individuals are still antibody-negative, especially in blood donors. Newly developed fourth-generation assays detect antibodies to HIV-1, including subtype O, and to HIV-2 and, simultaneously, p24 antigen of HIV-1. To evaluate this assay for daily routine work we compared it with different third-generation assays using sera from uninfected patients and patients with known HIV infection. The most interesting sera are those drawn during seroconversion from freshly infected patients. Whenever we encounter such a patient with acute HIV infection we store the serum in aliquots at -20 degrees C. Thus, we were able to establish our own seroconversion panel and use it in our laboratory for evaluation of new assays. The new test was shown to be able to detect all chronically HIV-infected individuals and four of six patients during seroconversion although in two of these patients conventional assays for HIV antibodies were still negative. The rate of unspecific reactivities was slightly higher as compared with third-generation assays.

Treatment guidelines are considered to be an important tool in steering patients to medical treatment. This study was conducted to analyze guidelines for the treatment of hepatitis C virus (HCV) infection in injection drug users (IDUs) in the European Union (EU) countries as a component of treatment access. National and international databases, expert contacts, professional societies, and health administrations were approached to acquire guidelines. According to their quality standard, guidelines were divided into expert opinions, semiofficial guidelines, official guidelines, and consensus processes. Recommendations for the treatment of HCV infection in IDUs vary substantially, from lack of recommendations and outright treatment disapproval to recommendations for treatment under specified circumstances. Recent guidelines that apply qualified process procedures that include literature research tend to be more permissive. Qualified guideline processes in each EU country and subsequently renewed pan-European guidelines are needed.

BACKGROUND: Risk factors for mother-to-child transmission of hepatitis C virus (HCV) are poorly quantified. METHODS: We conducted a European multicenter prospective study of HCV-infected pregnant women and their infants. Children with > or =2 positive HCV RNA polymerase chain reaction test results and/or anti-HCV antibodies after 18 months of age were considered to be infected. RESULTS: The overall HCV vertical transmission rate was 6.2% (95% confidence interval [CI], 5.0%-7.5%; 91/1479). Girls were twice as likely to be infected as boys (adjusted odds ratio [OR], 2.07 [95% CI, 1.23-3.48]; P=.006). There was no protective effect of elective cesarean section (CS) delivery on HCV vertical transmission (adjusted OR, 1.46 [95% CI, 0.86-2.48]; P=.16). HCV/human immunodeficiency virus-coinfected women more frequently transmitted HCV than did women with HCV infection only, although the difference was not statistically significant (adjusted OR, 1.82 [95% CI, 0.94-3.52]; P=.08). Maternal history of injection drug use, prematurity, and breast-feeding were not significantly associated with transmission. Transmission occurred more frequently from viremic women, but it also occurred from a few nonviremic women. CONCLUSIONS: Our results strongly suggest that women should neither be offered an elective CS nor be discouraged from breast-feeding on the basis of HCV infection alone. The sex association is an intriguing finding that probably reflects biological differences in susceptibility or response to infection.

Transfer of hepatitis B immunity occurs upon the transfer of immunologically active cells from the donor to the recipient by means of an organ graft. This has been repeatedly demonstrated for bone marrow and liver transplantations. Evidence is now presented for the transfer of anti-hepatitis B surface antibodies (anti-HBs) after kidney transplantation in rats. Kidney donors from one syngeneic and two allogeneic rat strains were immunized twice with 4 μg of recombinant hepatitis B vaccine. In week 6 after the first vaccination, kidney grafts were transplanted into Lewis (LEW) rats. Half of the recipients underwent daily immunosuppressive treatment with cyclosporin A (CsA). All recipients were vaccinated either after 10 weeks or 1 week postoperatively. Anti-HBs titer was measured weekly. Effective anti-HBs titers (10–227 mIU/ml, lasting for 1–7 weeks) were detected in 86% (25/29) of recipient rats, whose corresponding donors all had a titer above 15,000 mIU/ml. Immunosuppression enhanced the donor-derived immunity in terms of recipient-to-donor titer ratio, maximal titer and titer persistence.

Adoptive transfer of immunity to hepatitis B virus (HBV) is not provoked solely by bone marrow, but also by liver transplantation, although transiently. In the current study, simultaneous bone marrow transplantation, which possibly can increase the number of antibody-secreting cells, was performed to augment the efficacy of transferring HBV immunity. Stimulation of donor-derived immune cells by postoperative vaccination was used to investigate whether a secondary immune response can be induced in recipients. Orthotopic liver transplantation (n = 28), performed in three rat strain combinations representing different genetic constellations, was compared with bone marrow–augmented liver transplantation (n = 21). Donors had been vaccinated twice with recombinant hepatitis B surface antigen (HBsAg). Recipients surviving more than 10 weeks received a boost vaccination. All animals were monitored weekly for the presence of antibodies to HBsAg (anti-HBs). Effective anti-HBs titers were measured in 82% of liver recipients (23 of 28 recipients) and lasted from 2 to 9 weeks. Ninety percent of bone marrow–augmented liver recipients (19 of 21 recipients) seroconverted, with anti-HBs persisting from 2 to 12 weeks. A greater seroconversion rate, prolonged titer duration, and different pattern of titer development were observed in bone marrow–augmented liver recipients, although statistical significance could not be obtained because of the small numbers of comparable animals. Posttransplantation vaccination in recipients of combined grafts did not arouse a typical secondary antibody response, but showed a tendency toward an earlier and stronger response to vaccine in comparison to recipients without immune transfer. Simultaneous bone marrow transplantation showed an augmenting, but limited, effect on humoral immune transfer. Therefore, other potentially promising cellular strategies, such as transfer of in vivo and ex vivo stimulated antigen-specific cells should be pursued further. Improvement of the effect of postoperative vaccination possibly can be achieved by optimizing the immunization protocol. ( Liver Transpl 2002;8:397-404. )

Sera from 2,148 patients were tested with a third-generation microparticle enzyme immunoassay (MEIA), a confirmatory assay, and a reverse transcription-PCR. Overall, 85.6% of reactivities were confirmed, 13.2% were shown to be unspecifically reactive, and 1.2% were indeterminate. The rate of confirmed MEIA reactivities clearly depended on the strength of the reactivity.

We describe 2 patients who were initially positive for antibodies to hepatitis B surface antigen and who experienced a strong and sudden increase of hepatitis B virus (HBV) replication during highly active antiretroviral therapy (HAART). We found that reactivation of HBV replication during HAART can occur independently of lamivudine resistance or withdrawal of lamivudine, and in spite of increasing CD4(+) cell counts.

To investigate the vertical transmission of the newly described TT virus (TTV), serum and breast milk samples from 46 women as well as sera from their 47 newborns were examined for the presence of TTV DNA by PCR. TTV DNA was detected in 47.8% ( n = 22) of the women. All but one child born to these women were also viremic for TTV from the first sample onward. TTV DNA was found in 73.9% ( n = 17) of the breast milk samples derived from TTV viremic mothers. The one TTV-negative child born to a viremic mother remained negative during follow-up, although it was breast-fed. Our data show that TTV is highly effectively transmitted from mothers to their children during pregnancy. Although the majority of breast milk samples from viremic mothers are positive by TTV PCR, there is no need to discourage women from breast-feeding, because most children are TTV viremic even before breast-feeding begins.

Background: It is still unclear how many patients with hepatitis C virus (HCV) antibodies have viremia and hence are infectious. Objectives: To determine the chronicity of HCV infection by correlation of HCV antibodies with presence of viremia in long-term follow-up. Study design: In a longitudinal study sera of 4110 patients were analyzed with second generation HCV-enzyme immunoassay (EIA) and polymerase chain reaction (PCR). Only those patients were included in this study in whom sequential serum samples over a period of 2 years were available. To avoid preanalytical and analytical failures, we used a transport solution to prevent RNA degradation and a four-antigen recombinant immunoblot assay, established in our laboratory, for confirmation of antibody reactivity. Results: Of 2815 patients with confirmed HCV antibodies 2784 (98.9%) were also positive in HCV-PCR assay. False reactive EIA results were detected in 177 (13.7%) individuals as shown by confirmatory assay and PCR. Only one patient (0.04%) spontaneously lost detectable HCV viremia and subsequently HCV-specific antibodies. Conclusions: Our study clearly demonstrates that presence of confirmed HCV-specific antibodies correlates significantly (98.9%; P

Bei dialysepflichtigen Patienten mit chronischem Nierenversagen kommt es haufig zur parenteralen Transmission von Hepatitis B Infektionen [1]. Die prophylaktische Impfung scheitert an der schlechten Ansprechrate auf das Impfantigen, ein generelles Problem chronisch Kranker. Bei niereninsuffizienten Patienten fuhrt die Vakzination mit rekombinantem Impfstoff zu einer 50–80%igen Serokonversionsrate [2]. Patienten, die aufgrund ihrer Niereninsuffizienz transplantiert werden, haben ein noch hoheres Risiko einer Hepatitis B-Infektion [3] Ausserdem besteht die Gefahr der Reaktivierung einer durchgemachten Hepatitis B [4]. Die Therapie der Virusinfektion nach Transplantation gestaltet sich schwieriger als beim normalen Patienten, da die therapeutische Gabe von Interferon bei Transplantationsempfangern aufgrund des erhohten Risikos der Induktion eines Abstosungsreaktion nicht moglich ist [5].

DNA preparations from three different groups of serum samples were examined for HBV-DNA via a nested polymerase chain reaction assay (lower detection limit: 10 viral genomes in 100 μl serum): Group I consisted of 11 uninfected control sera, group II consisted of sera obtained from 11 HBV infected patients and group III consisted of 21 isolated anti-HBc positive samples. The 21 samples from group III were HBV-DNA negative according to a conventional non-nested PCR assay and hybridization with a 32P-labelled probe. Using nested PCR and mass spectrometry, HBV-DNA was detected in none of group I and in all of group II samples. In 11 out of 21 (52%) of the isolated anti-HBc positive sera from group III, HBV-DNA was detected. No correlation was observed between HBV-DNA positivity and anti-HBc titers. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided a fast, sensitive and non-radioactive assay for the detection of PCR products without the need for gel electrophoresis or hybridization with labelled probes.

Rhodococcus equi is a rare but well-documented cause of cavitary pneumonia in immunocompromised patients. In this report the first case of R. equi infection manifesting as vertebral osteomyelitis is described. A 39-year-old liver transplant recipient presented with recurrent pneumonia and a pleura-based lung abscess and subsequently developed osteomyelitis of the lower thoracic spine. Surgical debridement and prolonged treatment with rifabutin and clarithromycin resulted in clinical cure. In the literature, 12 other cases of R. equi infection in solid-organ transplant recipients have been reported. Ten of these patients had documented pulmonary disease and seven had extrapulmonary manifestations. Prolonged antibiotic therapy and surgical drainage resulted in clinical improvement in u90% of the reported cases.

The serodiagnosis of hepatitis C virus (HCV) infection was analyzed by a recombinant immunoblot assay (RIBA) with recombinant proteins encoded by the viral RNA isolated from our patients in Hamburg, Germany. The HCV RNA was amplified by PCR, and proteins encoded by the viral core and the NS3, NS4, and NS5 regions were expressed subsequently in Escherichia coli. The results obtained with our UKE RIBA were compared with the results of the Abbott HCV second-generation enzyme immunoassay (EIA). Serum samples from 270 patients, which were sent to us on the suspicion of HCV hepatitis and which were negative for hepatitis A virus and hepatitis B virus antibodies, were examined. In 227 cases (84.1%), there were identical positive (204 cases, 75.6%) or negative (23 cases, 8.5%) results in both tests. In 32 cases (11.9%), the reactive Abbott second-generation HCV EIA results could not be confirmed by the UKE RIBA and the HCV PCR. In follow-up studies conducted over 1 year, these results did not change. In three cases (1.1%), the UKE RIBA presented a positive result while the Abbott second-generation HCV EIA was negative. All three cases were positive in the HCV PCR and showed seroconversion in an HCV EIA 4 to 6 weeks later. In addition, 33 patient serum samples were examined by UKE RIBA in parallel with the Ortho RIBA 2.0. In three cases (9.1%), a positive Ortho RIBA 2.0 result could not be confirmed by the UKE RIBA and the HCV PCR. All three patients were free of complaints. The UKE RIBA showed also a smaller number of indeterminate results (3.0%) than the Ortho RIBA 2.0 (24.2%). This comparison study demonstrates that the commercially available HCV antibody tests should be further improved.

Spinal cord chimeras were produced by replacing a small fragment of neural tube of a 2-day-old White Leghorn chicken embryo with a similar fragment from a Japanese quail embryo. The embryo mortality w

Needle sharing among drug addicts leads to the transmission of infectious diseases such as hepatitis B and AIDS. After development of a test system based on gene technology against the hepatitis C virus (HCV), drug addicts have been regarded as an important reservoir for hepatitis C. In our study 113 (40.1%) out of 282 addicts who died from drug abuse in Hamburg between 1988 and 1990 had antibodies against HCV (anti-HCV). The prevalence of anti-HCV differed in various age groups; the highest prevalence was found in addicts aged 30-34 years. Co-infections with hepatitis B and hepatitis C virus were found in 57 drug addicts (59.4%) out of 96 deceased with antibodies to hepatitis B (anti-HBc), whereas only 8 out of 23 HIV-infected were anti-HCV positive (34.8%).

The prevalence of hepatitis G virus (HGV) and hepatitis C virus (HCV) infection was determined by reverse transcription-PCR in 777 individuals with and without risk factors for viral transmission via blood. From our results we conclude that transmission of HGV and that of HCV are favored by similar risk factors.

Plasma, tear fluid and swabs from eye, nose and pharynx of 33 patients were examined for presence of hepatitis C virus (HCV) RNA by polymerase chain reaction (PCR). All samples from plasma, tear fluid and eyeswabs were found to show a positive reaction in HCV-RNA PCR. Remarkably, we regularly found greater amounts of amplification products in tear fluid and eyeswabs compared to plasma using the same conditions for sample preparation.

In 1989 blood samples from 2581 fatalities investigated at the Institute of Forensic Medicine in Hamburg were screened for HIV-1-antibodies. Sera from 13 corpses were confirmed positive for HIV-1 (prevalence rate approx 0.5 %).-Viable HIV was found in blood cultures of 4 cadavers stored under non-refrigerated conditions up to 36 hours after death.

Bovine hepacivirus (BovHepV) is a recently added member to the growing genus Hepacivirus within the family Flaviviridae. Animal hepaciviruses are rarely characterized so far. Apart from norway rat hepacivirus which represents a promising HCV surrogate model, only equine hepaciviruses have been studied to some extent. BovHepV has been initially identified in bovine samples and was shown to establish persistent infections in cattle. However, consequences of those chronic infections, humoral immune response and the possibility of an extended host spectrum have not been explored so far. Therefore, we here investigated (a) the presence of anti-NS3-antibodies and viral RNA in cattle herds in Germany, (b) the course of infection in cattle, and (c) the host tropism including zoonotic potential of bovine hepaciviruses. Our results show that 19.9% of investigated bovine serum samples had antibodies against BovHepV. In 8.2% of investigated samples, viral RNA was detected. Subsequent genetic analysis revealed a novel genetic cluster of BovHepV variants. For 25 selected cattle in a BovHepV positive herd the presence of viral genomic RNA was monitored over one year in two to three months intervals by RT-PCR in order to discriminate acute versus persistent infection. In persistently infected animals, no serum antibodies were detected. Biochemical analyses could not establish a link between BovHepV infection and liver injury. Apart from a single sample of a pig providing a positive reaction in the antibody test, neither BovHepV-specific antibodies nor viral RNA were detected in porcine, equine or human samples implying a strict host specificity of BovHepV.