[Risk estimation of blood-borne infections by emergency room personnel]. / Risikoeinschätzung von blutübertragbaren Infektionen durch die Schockraummitarbeiter.

BACKGROUND: Emergency department personnel are at risk of occupational exposure to blood-borne pathogens. Previous studies have shown that the prevalence of human immunodeficiency virus (HIV), hepatitis B (HBV) and hepatitis C (HCV) virus infections among trauma patients is higher compared to the general population. OBJECTIVES: The aim of the study was to investigate the compliance rates of trauma team members in applying standard precautions, knowledge about the transmission risk of blood-borne infections and perceived risk of acquiring HIV, HBV and HCV. METHODS: An anonymous questionnaire was distributed to 100 trauma team members including physicians, nurses and medical students from different medical departments (e.g. surgery, radiology, anesthesia and internal medicine). RESULTS: The results of the questionnaire showed that trauma team members had insufficient knowledge of the risk of blood-borne pathogens, overestimated the risk of HCV infection and underused standard precautions during treatment of emergency trauma patients. CONCLUSION: Further educational measures for emergency department personnel are required to increase the knowledge of occupational infections and compliance with standard precautions. Every healthcare worker needs to be sufficiently vaccinated against HBV. In the case of injury awareness of all measures of post-exposure prophylaxis is of utmost importance for affected personnel.

[Prevention of nosocomial transmission of human immunodeficiency virus (HIV) from HIV-positive healthcare workers. Recommendations of the German Association for the Control of Viral Diseases (DVV) e.V. and the Society for Virology (GfV) e.V]. / Prävention der nosokomialen Übertragung von humanem Immunschwächevirus (HIV) durch HIV-positive Mitarbeiterinnen und Mitarbeiter im Gesundheitswesen. Empfehlungen der Deutschen Vereinigung zur Bekämpfung der Viruskrankheiten (DVV) e.V. und der Gesellschaft für Virologie (GfV) e.V.

To the best of our knowledge, the German Association for the Control of Viral Diseases (DVV) e.V. and the Society for Virology (GfV) e.V. are the first in Europe to provide precise recommendations for the management of health care workers (HCWs) who are infected with human immunodeficiency virus (HIV). Requirements for HIV-infected HCWs need to be clearly defined. With a permanent viral burden of less than or equal to 50 copies/mL, HIV-positive HCWs are allowed to perform any surgery and any invasive procedure, as long as the infected HCW uses double-gloving, undergoes follow-up routinely by occupational medicine professionals, undergoes a quarterly examination of viral burden, and has a regular medical examination by a physician who has expertise in the management of HIV. Unrestricted professional activity is only possible with a strict compliance to take antiretroviral therapy and if the HIV-infected HCW strictly adheres to the recommended infection control procedures. Complete compliance with the recommendation almost certainly leads to no HIV transmission risk in patient care.

[Blood-borne infections and the pregnant health care worker. Risks and preventive measures]. / Blutübertragbare Infektionen und die schwangere Mitarbeiterin im Gesundheitswesen. Risiko und Präventionsmaßnahmen.

Due to the increasing proportion of women in health care, as well as changes in working conditions (implementation of safety devices, minimally invasive/endoscopic procedures) the question arises whether the applicable laws and regulations for the protection of working mothers are outdated and should be updated.Individual risk analysis, as well as the inclusion of the pregnant health care worker in the decision-making process with regard to continuation or modification of the work practice serves as a protection of the expectant mother and unborn child and allows a continuation of the occupational activities.

Infection of cultured intestinal epithelial cells with severe acute respiratory syndrome coronavirus.

To identify a model for the study of intestinal pathogenesis of severe acute respiratory syndrome (SARS) we tested the sensitivity of six human intestinal epithelial cell lines to infection with SARS coronavirus (SARS-CoV). In permissive cell lines, effects of SARS-CoV on cellular gene expression were analysed using high-density oligonucleotide arrays. Caco-2 and CL-14 cell lines were found to be highly permissive to SARS-CoV, due to the presence of angiotensin-converting enzyme 2 as a functional receptor. In both cell lines, SARS-CoV infection deregulated expression of cellular genes which may be important for the intestinal pathogenesis of SARS.

Acute retinal necrosis six years after herpes simplex encephalitis: an elusive immune deficit suggested by insufficient test sensitivity.

A patient presented with acute retinal necrosis of the left eye. Demonstration of herpes simplex virus (HSV) DNA in the aqueous humour confirmed the diagnosis. Negative results of HSV type-specific antibody tests based on gG antigens suggested a primary HSV infection. However, the patient had a past history of laboratory-confirmed herpes simplex encephalitis 6 years ago. Using antibody tests based on whole viral lysate antigens, he was seropositive from the onset, and immunoblot testing confirmed a lack of anti-gG reactivity. To be able to assess whether this might be related to the apparent inability of his immune system to suppress clinically symptomatic HSV infection, serial samples were tested by an HSV neutralisation test and a whole-blood flow cytometric assay to determine the frequency of HSV-specific CD4 lymphocytes. However, this did not yield evidence of obvious immunodeficiency; the patient reacted similarly to known positive controls by both assays. Although type-specific HSV serological tests based on gG are generally more specific than those based on whole viral lysate antigens, they have a somewhat lower sensitivity, as a certain percentage of HSV-infected individuals do not develop antibodies against gG, and others may suffer a secondary loss of anti-gG reactivity. Thus there is a risk of missing individual infected patients. Unless this potential problem is recognised, serious consequences might possibly result. We therefore urge virologists and clinicians to exercise great care if highly specific antibody assays based on recombinant proteins are employed.

Prions and orthopedic surgery.

Prions are a novel class of infectious agents that cause subacute encephalopathy in man and animals as human Creutzfeldt-Jakob disease (CJD), sheep scrapie and bovine spongiform encephalopathy (BSE). Previously, prions were shown to be transmitted by neuro- and ophthalmosurgical measures and by application of brain-derived therapeutic hormones. Recently, prions have been detected in blood specimens of experimentally infected monkeys indicating a principal threat to transfusion medicine, furthermore in human or bovine materials used in reconstructive surgery. In this article the risk of prion transmission from the surgeon to the patient or vice versa during (orthopedic) surgery is reevaluated including the issues of blood transfusion. This is accomplished based on recent epidemiologic findings and biometric calculations on the spread of prions in animals and humans as well as in terms of experimental data on artificially contaminated medical materials and devices. The overall risk of prion transmission in orthopedic surgery is considered very low if adequately prepared and sterilized materials and devices are used.

Polyomavirus viruria in bone marrow transplant recipients: lack of correlation with clinical symptoms.

BACKGROUND: Human polyomavirus (HPV) infection is controversially discussed as a factor influencing the outcome of bone marrow transplantation (BMT). PATIENTS AND METHODS: Here we report on 62 patients undergoing BMT with clinical signs of urocystitis, such as micro- or macrohematuria with more or less severe dysuria. These patients were tested for the presence of HPV in urine specimens (n = 80) by transmission electron microscopy (TEM). RESULTS: HPV viruria was found in 35 patients (56%); 33 (94%) of them had hemorrhagic cystitis, whereas two (6%) were suffering from dysuria only. Among the patients with hemorrhagic cystitis, ten (30%) presented with macrohematuria, four (12%) with microhematuria and 19 (58%) with micro- or macrohematuria and severe dysuria. 26 of 27 HPV-negative patients (96%) showed hemorrhagic cystitis with either macrohematuria (n = 7) (26%) or microhematuria (n = 15) (56%). Four patients (15%) suffered from hematuria and dysuria, one patient (4%) from dysuria only. CONCLUSION: Although HPV-negative patients tended to present with less severe clinical symptoms, overall no statistically significant influence on the outcome of BMT was seen.

An epidemiological study of HIV-infections in Frankfurt/Main and other major cities in Germany.

Epidemiological studies can help to understand the effects of medical treatment of HIV infections. Accordingly, this study was designed to discuss the most important parameters in Frankfurt/Main and other big German cities from 1984 to 2000. The number of HIV tests performed by Frankfurt's Virology has been decreasing continually since 1991. A decrease of new infections in men could be registered, whereas the number of HIV infected women rose. From 1985 to 2000 an annual mean value of 478 HIV infected men and 121 HIV infected women was registered in Frankfurt. The gender proportion was followed up for Frankfurt and Hamburg since 1985, for Berlin, Munich, and Cologne since 1993. All but one city showed a significant decline of infected males, only Berlin did not show any obvious changes in this proportion. Over the last twelve years the average age of men and women tested positive for the first time increased. An obvious shift could be discerned during the last two years concerning the distribution of risk groups. The percentage of HIV infected homosexuals and female i.v. drug addicts sank significantly over the last two years, the number of women infected by heterosexual contacts is still increasing when compared to data compiled from 1988 to 1992, and varies between 44% and 33%. During the same time-span a significant shift in first onset of AIDS-defining illnesses was observed. PCP (pneumocystis carinii pneumonia)--formerly represented with 35.5%--decreased and is now surpassed by tuberculosis with 25.5%. The general gender proportion (3:1) could not be reflected by AIDS-defining diseases of which NHL (non-Hodgkin-lymphoma) seems to have the shortest time-span (6.5 months) between the occurrence of illness and death.

AIDS-related body cavity-based lymphoma. A case report.

BACKGROUND: Body cavity-based lymphomas are rare malignancies in human immunodeficiency virus (HIV)-infected patients, but because of their unusual clinical, morphologic and immunophenotypic features, they are recognized as a distinct subgroup of lymphomas connected to human herpesvirus 8 (HHV-8) infection. CASE: A 39-year-old, HIV-positive, homosexual man was admitted to the hospital because of a left-sided pleural effusion that contained malignant lymphoid cells. He responded partially to a low-dose cyclophosphamide/doxorubycin/vincristine/prednisone regimen and died five months after the diagnosis of lymphoma. On cytology, the sediments contained exclusively large, round, neoplastic, lymphoid cells with abundant basophilic cytoplasm and large, round nuclei with prominent nucleoli. Many cells had immunoblastic features, and some had plasmocytoid differentiation. Mitotic figures were numerous. On flow cytometry, the homogeneous population of large cells expressed CD45, CD38, HLA-DR and CD7 positivity. Other specific T-, B- and NK-cell markers tested negative. Polymerase chain reaction demonstrated Epstein-Barr virus (EBV) and HHV-8 in the malignant effusion. CONCLUSION: Primary effusion from lymphoma with molecular evidence of HHV-8 and EBV coinfection represents a distinct clinical and morphologic entity in AIDS patients. However, immunophenotypic markers of malignant clones can be diverse in different cases.

Evaluation of diagnostic methods for the detection of cytomegalovirus in recipients of allogeneic stem cell transplants.

BACKGROUND: Although several diagnostic methods are available for the surveillance of patients at risk of human cytomegalovirus (CMV) infection and disease, little data is available on their comparative performances in the diagnostic setting. OBJECTIVES: To compare different assays for CMV detection, especially assays based on (quantitative) DNA and mRNA detection. STUDY DESIGN: Eight allogeneic bone marrow and stem cell transplant recipients at high risk for developing CMV disease (donor CMV-negative, recipient positive) were regularly tested for 7-20 weeks post-transplant by spin-amplification rapid culture from urine (viruria), antigenemia (pp65 assay), pp67 mRNA in whole blood (NASBA), and CMV DNA both qualitatively (in-house PCR, whole blood) and quantitatively (in-house PCR, plasma; Cobas Amplicor CMV Monitor Test, plasma and whole blood; Hybrid Capture, whole blood). RESULTS: Four patients (50%) suffered CMV reactivation during follow-up. Out of 104 sample dates, 41 (39.4%) yielded a positive CMV result in at least one assay. Out of the 28 samples tested by all assays, the highest percentage of positive results was obtained with the in-house quantitative PCR (60.7%), followed by the Hybrid Capture system (39.3%), the Cobas Amplicor CMV Monitor Test, plasma version (35.7%), the Cobas Amplicor CMV Monitor Test, whole blood version (32.1%), in-house qualitative PCR (28.6%), and the mRNA assay (21.4%). Viruria was positive in one sample and pp65 antigenemia was found in two samples. CONCLUSIONS: Despite a considerable incidence of CMV reactivations, pre-emptive anti-CMV chemotherapy prevented the development of CMV disease with the exception of one case. The molecular assays had superior sensitivity to conventional ones. The antigenemia assay proved unsuitable for the surveillance of hematological transplant patients. However, none of the tests recognized all timepoints with CMV reactivation. Further comparative studies are needed to determine their respective diagnostic values.

Kaposi's sarcoma-associated herpesvirus seroprevalence in selected german patients: evaluation by different test systems.

A total of 603 serum samples obtained from 12 different patient and control groups, including potentially cross-reactive sera, were tested for the presence of antibodies against Kaposi's sarcoma-associated herpesvirus (KSHV, HHV-8). The assays used were an inhouse immunofluorescence test (IFT) employing latent KSHV antigens and a prototype enzyme-linked immunosorbent assay (ELISA) coated with recombinant latency-associated KSHV nuclear antigen (LANA, open reading frame 73) and the K8.1 protein. Sera giving discrepant results were additionally tested with two commercial IFTs employing KSHV latent and lytic antigens, respectively. The low KSHV seroprevalence rate found in blood donors (3.0% by in-house IFT, 2.0% by recombinant ELISA) was comparable to that found previously in Western European countries. The highest KSHV seroprevalence rates were found in patients with Kaposi's sarcoma (100% by both assays), followed by HIV-infected men without Kaposi's sarcoma (23.3% by in-house IFT and 17.8% by ELISA) and women (15.7% by in-house IFT and 13.7% by ELISA). Overall correlation between both assays was 91.2%, with the highest rate of discordant results occurring in HIV-infected male subjects. Retesting of the 53 discrepant samples by the commercial IFTs revealed the best, albeit low, correlation between the in-house IFT and the commercial latent antigen IFT and poor correlations between the other assays. Apart from patients with autoimmune antibodies, there was no significant degree of non-specific reactivity in either of the KSHV tests due to antibodies against Epstein-Barr virus and human herpesvirus 6. Despite the lack of a "gold standard" for KSHV antibody detection, the fact that the results obtained overall agreed rather well indicates their suitability for conducting seroepidemiological studies.

High frequency of HCV infection in individuals with isolated antibody to hepatitis B core antigen.

Although isolated antibody to hepatitis B core antigen (anti-HBc) is frequently nonspecific or may be the only serological marker of past self-limiting hepatitis B, where antibodies against the surface antigen have disappeared, isolated anti-HBc seropositivity is frequently associated with chronic hepatitis B in HIV- and HCV-infected individuals. Of 5,520 samples that tested positive for anti-HBc (IMx and AxSYM CORE, Abbott, Delkenheim, Germany) at the Institute of Virology, University Clinic Frankfurt during the time interval from January 1994 to February 1996, 643 (11.6%) were isolated anti-HBc-reactive in the IMx and AxSYM CORE assays (inhibition values >90%). There was a statistically significant association between isolated anti-HBc seropositivity and HCV and HIV/HCV coinfection (p < 0.05). A total of 190 samples were available for further testing. Six (3.2%) of 190 isolated anti-HBc-positive samples were considered false-positive since they were only positive in the AxSYM or IMx CORE assay and a linear decrease of the measured signal could not be observed in dilution series. Of 184 serum samples tested with nested PCR using primers of the S genome region, only 6 (3.3%) were HBV DNA-positive. Anti-HBc-IgM antibody could be detected in 3 (1.6 %) of the tested samples using the IMx CORE-M. With the more sensitive VIDAS HBc IgM specific IgM antibody was detected in 15 (8.5%) of 177 samples at concentrations ranging from 10 to >200 Paul Ehrlich Institute U/ml. HIV or HCV coinfection was present in 28.1% and 37.5% of isolated anti-HBc-positive individuals, respectively. We conclude from our observations that only a limited proportion of anti-HBc-isolated individuals are potentially infectious, however anti-HBc-IgM which is detectable in any form of liver disease associated with HBV infection was present in more than 8% of the individuals. Of isolated anti-HBc-positive sera 37% were positive for anti-HCV, suggesting that anti-HCV antibody testing should be performed in isolated anti-HBc-positive individuals.

Aphidicolin glycinate inhibits human neuroblastoma cell growth in vivo.

Aphidicolin is a fungal derived tetracyclic diterpene antibiotic. It is selectively toxic for neuroblastoma (NB) cells in vitro but has no significant effects on the viability of normal human cells and a variety of other tumor entities. We evaluated the antitumoral effects of the water soluble ester aphidicolin glycinate (AphiG) on established human NB xenografts from UKF-NB-3 cells in athymic (nude) mice. Furthermore, we explored the efficacy of direct intraneoplastic and systemic delivery of AphiG. Systemic administration of AphiG (60 mg/kg intraperitoneally, twice per day on 10 consecutive days) significantly suppressed tumor growth but was not able to induce any cures. In contrast, intratumoral AphiG injections (60 or 40 mg/kg/twice a day for 4 days) induced complete tumor regression. Two weeks after the end of treatment no tumor cells were microscopically detectable. Animals were free of tumor for more than 90 days. Histologic examination of inner organs and bone marrow did not reveal any apparent toxic effects of AphiG. These data strongly indicate that AphiG deserves further evaluation as a specific treatment for neuroblastoma.

Application of automated thermal disinfection instead of sterilisation procedures for treatment of rotating dental instruments: efficacy against viruses?

In dentistry it is of primary importance to take into consideration microbial transfer due to the nature of the construction of rotating dental instruments. This aspect was the starting point for our research with the question whether or not sterilisation is fundamentally necessary for slow and high speed hand pieces to make them "safe" out of a virological point of view, or whether a thermal disinfection could also possibly be adequate for this purpose. In this context, we tested the efficiency of the cleaning and disinfection capacity of an automated steam disinfection and sterilisation unit (Sirona Hygiene Center, Siemens, AG, Bensheim) intended to the hygienic treatment of dental instruments with respect to viruses. In model tests the corresponding instruments were experimentally infected with herpes simplex virus type 1 (HSV) and simian vacuoling virus (SV40). As indicator systems we used for both cell cultures (measurement of the degree of infectiosity) and (for HSV) polymerase chain reactions (PCR; determination of viral nucleic acids). In the tests for (residual) infectiosity after thermal disinfection (as an isolated step of the Hygienic Centre) and also for a combination of cleaning and subsequent thermal disinfection (also after protein application), no infectious virus could be found in the interior of the slow handpieces and turbines tested. In opposite to this, infectious HSV and SV40 could be found after completion of every isolated cleaning program in the turbine (in all three ducts) and in the slow handpiece (only in the gearbox duct in the case of HSV, and in the case of SV40 also in the water and air ducts in very small amounts). The PCR analyses showed that no nucleic acids could be found in both instruments (in the air and water ducts) following a practice-relevant combination of cleaning and disinfection, but that PCR-positive signals were obtained for the larger-volume gearbox and drive and return air ducts in 1 or 2 of 3 test samples. The detection of viral nucleic acid proves that it is not a matter of complete removal of the infectious agents from the instruments, but rather that they are simply inactivated. Since the test samples did not show similar amounts of viral nucleic acid, it demonstrates the strong influence of the basic parameters (efficiency of the virus contamination, flushing conditions and the like). Our experimental model demonstrate that thermal disinfection may be adequate to prevent virus contamination of rotating dental instruments, while compulsory sterilisation is not mandatory. Further studies are necessary to demonstrate whether the present data are applicable for rotating dental instruments from other manufactures.