Viral determinants predicting hepatitis B surface antigen (HBsAg) seroclearance in HIV-/HBV-coinfected patients.

The aim of this retrospective study was the identification of clinically useful viral determinants for the prediction of hepatitis B surface antigen (HBsAg) seroclearance and sustained virological response in hepatitis B virus/human immunodeficiency virus (HBV-/HIV)-coinfected patients receiving HBV-active combined antiretroviral therapy (cART). Quantification of HBsAg, HBeAg and HBV DNA before and after initiation of HBV-active cART in a cohort of 59 HIV-/HBV-coinfected patients was performed. Calculations of receiver operating characteristics (ROC) and Kaplan-Meier analysis were used for the identification of predictors of HBsAg seroclearance for HBeAg-positive [HBeAg(+); n = 36] and HBeAg-negative [HBeAg(-);n = 23] patients. HBeAg(+) patients with an HBsAg on-treatment decline ≥ 1 log IU/mL per year achieved higher HBsAg loss rates (P = 0.0294), whereas the quantification of HBeAg had no predictive value for HBsAg seroclearance. Among HBeAg(-) patients, a pretreatment baseline cut-off level of HBsAg ≤ 100 IU/mL was highly predictive for HBsAg seroclearance. No significant influence of the HBV genotype on HBsAg seroclearance was observed among the entire cohort. Quantitative determination of HBsAg provides a clinically useful viral parameter for the prediction of HBsAg seroclearance both in HBeAg(+) and HBeAg(-) HIV-/HBV-coinfected patients receiving HBV-active cART.

Rapid identification of neuraminidase inhibitor resistance mutations in seasonal influenza virus A(H1N1), A(H1N1)2009, and A(H3N2) subtypes by melting point analysis.

The high mutation rate of influenza virus, combined with the increasing worldwide use of influenza virus-specific drugs, allows the selection of viruses that are resistant to the currently available antiviral medications. Therefore, reliable tests for the rapid detection of drug-resistant influenza virus strains are required. We evaluated the use of a procedure involving real-time polymerase chain reaction (PCR) followed by melting point analysis (MPA) of hybrids formed between the PCR product and a specific oligonucleotide probe for the identification of point mutations in the influenza A virus neuraminidase gene (NA) that are associated with oseltamivir resistance [resulting in the amino acid change H275Y for seasonal and pandemic influenza A(H1N1) viruses and E119V for A(H3N2) viruses]. Therefore, 54 seasonal A(H1N1) (12 oseltamivir-resistant and 42 sensitive strains), 222 A(H1N1)2009 (5 resistant, 217 sensitive), and 51 A(H3N2) viruses (2 resistant, 49 sensitive) were tested by MPA, and the results were compared to those obtained by sequencing the NA gene. The results clearly indicate that the identification of drug resistance mutations by MPA is as accurate as sequencing, irrespective of whether MPA is performed using clinical material or the corresponding isolate. MPA enables a clear identification of mutations associated with antiviral resistance.

Antibodies induced with recombinant VP1 from human rhinovirus exhibit cross-neutralisation.

Human rhinoviruses (HRVs) are the major cause of the common cold and account for 30-50% of all acute respiratory illnesses. Although HRV infections are usually harmless and invade only the upper respiratory tract, several studies demonstrate that HRV is involved in the exacerbation of asthma. VP1 is one of the surface-exposed proteins of the viral capsid that is important for the binding of rhinoviruses to the corresponding receptors on human cells. Here we investigated its potential usefulness for vaccination against the common cold. We expressed VP1 proteins from two distantly related HRV strains, HRV89 and HRV14, in Escherichia coli. Mice and rabbits were immunised with the purified recombinant proteins. The induced antibodies reacted with natural VP1 and with whole virus particles as shown by immunoblotting and immunogold electron microscopy. They exhibited strong cross-neutralising activity for different HRV strains. Therefore, recombinant VP1 may be considered a candidate HRV vaccine to prevent HRV-induced asthma exacerbations.

Relative seroprevalence of human herpes viruses in patients with chronic lymphocytic leukaemia.

BACKGROUND: Herpes virus infections may have a significant role in chronic lymphocytic leukaemia (CLL) due to their ability to modulate the host's immune system. MATERIALS AND METHODS: We examined the seroprevalence of four herpes viruses [Cytomegalovirus (CMV), Epstein-Barr Virus (EBV), human herpes virus (HHV)-6 and -7] in a cohort of European CLL patients (cohort 1, n = 100) in relation to the immunoglobulin variable heavy (IGHV) chain gene use and compared serological results with those obtained from age- and gender-matched healthy adults (n = 100). RESULTS: CMV-seroprevalence was significantly higher in CLL cohort 1 (79%) than in the control cohort (57%, P = 0.001); the seroprevalence of EBV (89% vs. 94%), HHV-6 (73% vs. 60%), or HHV-7 (35% vs. 35%) was not. In CLL cohort 1, use of IGHV3-30 was more prevalent among CMV-seropositive and of IGHV3-21 among HHV-7-seronegative cases. To investigate the generalizability of these findings, we investigated the herpes virus seroprevalence in a second cohort of age-matched CLL patients from a different geographical area (USA, n = 100, cohort 2). In cohort 2, CMV-seroprevalence was comparable with that of the control cohort (53%). Seroprevalence of EBV, HHV-6 and HHV-7 were 85%, 88% and 73% respectively. In CLL cohort 2, use of IGHV3-30 or IGHV3-21 was not associated with any of the herpes viruses investigated. CONCLUSIONS: CMV-seropositivity is associated with CLL in selected patient cohorts. However, the considerable variation in herpes virus-specific seropositivity between geographically distinct CLL cohorts indicates that seropositivity for any of the four human herpes viruses investigated is not generally associated with CLL.

A novel PCR-based point-of-care method facilitates rapid, efficient, and sensitive diagnosis of influenza virus infection.

OBJECTIVE: The aim of this single-centre study was the comparative analysis of the GeneXpert (Cepheid Inc.) and the LIAT (Roche) system for the rapid polymerase chain reaction (PCR)-based detection of influenza A (IA) and influenza B (IB) viruses. PATIENTS AND METHODS: During the 2017-2018 flu season, 651 prospectively collected samples (throat and nasal swabs) of patients with symptoms of influenza-like illness or acute respiratory infection were tested for the presence of IA and IB viruses using the GeneXpert and LIAT systems. To evaluate the usefulness for near-patient testing, a LIAT system was installed at the Department of Emergency Medicine, and sample testing was performed on site. Reference testing of all samples was performed with the Xpert Flu assay and for 313 samples in addition with the Xpert Xpress Flu/RSV (respiratory syncytial virus) assay at the central laboratory. Analysis of all samples was carried out within 24 hr after collection. RESULTS: Overall, 267 of the 651 samples analysed were positive for influenza viruses in at least one of the three assays investigated (IA, 88; IB, 179). The overall rates of agreement between the LIAT assay and the Xpert Flu assay was 96.0% for the detection of IA and IB viruses. The sensitivity and specificity of the LIAT assay compared to the Xpert Flu assay for the detection of IA was 98.80% (95% confidence interval (CI) 93.47-99.97%) and 99.12% (95% CI, 97.96% to 99.71%) and for the detection of IB 98.76% (95% CI 95.58-99.85%), and 96.33% (95% CI 94.26-97.81%), respectively. The LIAT assay showed a statistically significant higher detection rate of IB virus than the Xpert Flu assay (p <0.01). No significant difference was found between the detection rate of the LIAT assay and the Xpert Xpress Flu/RSV assay. The mean time to the availability of a definite test result was significantly shorter with the on-site LIAT system than the GeneXpert system (mean 59 min saving time; p <0.01). CONCLUSION: The LIAT system represents a robust and highly sensitive point-of-care device for the rapid PCR-based detection of influenza A and influenza B viruses.

Mumps outbreak in young adults following a festival in Austria, 2006.

Mumps is not a mandatorily notifiable disease in Austria. However, in the first week of May 2006, a sudden increase in serologically confirmed cases of mumps, confined to three public health districts of the southern Austrian province of Carinthia, was identified by the Austrian Reference laboratory for MMR. An epidemiological investigation of this cluster of mumps cases was performed. A total of 214 cases fulfilled the outbreak case definition; 143 cases were laboratory confirmed and 71 cases were epidemiologically linked and fulfilled the clinical picture of the case definition. The vaccination status was known for 169 patients. Nearly half of the cases for whom the vaccination status was known occurred in non-vaccinated persons, another 40% were vaccinated with one dose of the vaccine and 11% had received two doses. Only four mumps cases occurred in children aged 14 years or younger, indicating that the vaccination coverage and the acceptance of the recommended childhood vaccinations have strongly improved within the past 15 years. Vaccination scheme failure but not vaccine failure is primarily to blame for this mumps outbreak.

A human case of travel-related rabies in Austria, September 2004.

A young male Austrian tourist, aged 23 years and unvaccinated against rabies, was bitten by a dog in Morocco in July 2004. One month later he was hospitalised in Ceuta with symptoms compatible with rabies. He died on 23 September in an Austrian hospital after a diagnosis of rabies was confirmed by FAT, IHC and RT-PCR (including sequencing) of the neck skin and the RT-PCR (including sequencing) of the pharyngeal swab. This Austrian case of laboratory confirmed rabies highlights the urgent need for reinforcement of the international recommendations for travel vaccinations.

A human case of travel-related rabies in Austria, September 2004.

A young male Austrian tourist, aged 23 years and unvaccinated against rabies, was bitten by a dog in Morocco in July 2004. One month later he was hospitalised in Ceuta with symptoms compatible with rabies. He died on 23 September in an Austrian hospital after a diagnosis of rabies was confirmed by FAT, IHC and RT-PCR (including sequencing) of the neck skin and the RT-PCR (including sequencing) of the pharyngeal swab. This Austrian case of laboratory confirmed rabies highlights the urgent need for reinforcement of the international recommendations for travel vaccinations.

Endemic Kaposi's sarcoma in human immunodeficiency virus type 1-seronegative persons: demonstration of retrovirus-like particles in cutaneous lesions.

In 1984, Greek physicians reported on the clustering of cases of Kaposi's sarcoma (KS) on the Peloponnesus peninsula. To gain more insight into its pathogenesis, we studied the seroepidemiologic and clinicopathologic characteristics of 12 Greek KS patients (eight male/four female) five of whom were residents of an endemic area on the Peloponnesus. These patients were in good general health with ages ranging from 48 to 80 years, had no clinical signs of immunodeficiency, and combined the features of both classic and epidemic KS in that they displayed not only involvement of acral areas but also widespread mucocutaneous lesions. Routine laboratory data were within normal limits; no patient had HTLV-1 and HIV-1/2 antibodies, but all patients had antibodies to several herpesviruses. The histopathology was characteristic of KS with the peculiar feature of a dense infiltrate composed predominantly of CD4+ T lymphocytes. Immunoenzymatic/morphologic studies of the KS cells were consistent with their origin from lymphatic endothelium. Outstanding ultrastructural findings were tubuloreticular structures and cylindrical confronting cisternae, structures that are indicative of an ongoing viral infection. Indeed, extensive electronmicroscopic studies resulted in the detection of retrovirus-like particles in close association to KS cells in five of 12 patients. This in situ observation opens the possibility that this retro-virus contributes to KS development.

CMV-viraemia during allogenic bone marrow transplantation in paediatric patients: association with survival and graft-versus-host disease.

Sixty-one consecutive paediatric patients undergoing allogenic bone marrow transplantation (BMT) were screened prospectively for cytomegalovirus (CMV)-viraemia by PCR. Sixteen patients (26%) presented with single or recurrent CMV-viraemia between day -7 and + 100. Although only four of them had evidence of CMV-disease, there was a significant difference in the incidence of acute Graft versus Host Disease (GVHD) grade III-IV (75% vs 15.5%), liver-involvement (68% vs 13%) and the incidence of chronic GVHD (83% vs 13.8%) between CMV-PCR-positive and CMV-PCR-negative patients. Transplant-related mortality (TRM) was 43.7% in the CMV-PCR-positive group versus 13% in patients which had no evidence for CMV-viraemia. In all but one cases mortality in CMV-PCR positive patients was GVHD-associated. Pre-emptive therapy with gancyclovir in case of CMV-viraemia seemed to have no impact on incidence and severity of GVHD.

Prophylactic use of hyperimmunoglobulin for cytomegalovirus infection in heart transplantation.

Cytomegalovirus (CMV) infection in solid organ recipients can endanger the immunosuppressed patient and increase vulnerability to secondary infections and the high risk of rejection triggered by the viral disease. The effect of passive immunization against CMV was examined in 69 heart transplant patients. The patients received weekly administrations of 1 ml/kg of CMV hyperimmunoglobulin from the day of transplantation until the 30th postoperative day. Forty-four of the patients were monitored clinically and serologically up to the 120th postoperative day. Nine patients showed clinical and serologic signs of CMV infection; in 15 the only evidence of CMV infection was a rise in antibody titers. The remaining 20 patients showed no clinical or serologic signs of CMV infection. Three patients who were seronegative preoperatively remained seronegative until the end of the observation period. The results indicate a potential therapeutic benefit of hyperimmunoglobulin prophylaxis to prevent infectious complications due to CMV in heart transplant patients.

[Immunoglobulin A serum antibodies against the capsid antigen of Epstein-Barr virus in the differential diagnosis and follow-up of nasopharyngeal cancer]. / Immunglobulin-A-Serumantikörper gegen das Kapsidantigen des Epstein-Barr-Virus zur Differentialdiagnose und Verlaufskontrolle des Nasopharynxkarzinoms.

IgA antibodies to Epstein-Barr virus capsid antigen (IgA anti-VCA) can be detected in sera of patients with certain types of nasopharyngeal carcinoma (NPC). IgA anti-VCA titres were determined by the indirect immunofluorescence technique. 17 control patients with benign diseases or carcinomas of the head and neck other than NPC had negative IgA anti-VCA titres less than or equal to 1:16. NPC was diagnosed histologically according to the Cologne modification of the WHO classification. Among 16 cases of untreated or recurrent NPC, a rare disease in Europe, seen over the past three years, those with undifferentiated carcinomas with and without lymphoid stroma and the non-keratinizing carcinomas with lymphoid stroma were IgA anti-VCA positive (1:32 to 1:512), whereas patients with squamous cell carcinomas were negative. In four cases the primary tumour had not been diagnosed by other ENT doctors in spite of known regional or distant metastases consisting of undifferentiated carcinomas with or without lymphoid stroma. IgA anti-VCA antibodies in the sera of these patients indicated the probable site of the primary tumour. NPC was verified by biopsy in all these cases. In 2 serologically negative patients the original diagnosis of undifferentiated NPC with lymphoid stroma had to be revised to malignant Non-Hodgkin lymphoma. In the follow-up of 6 NPC patients the trend of changes in IgA anti-VCA titres correlated with the course of the disease, but the minute tumour-related changes could be detected only when at least two previous sera of the same patient were included in every test.(ABSTRACT TRUNCATED AT 250 WORDS)

[The 1984/85 rubella epidemic in Austria]. / Die Rötelnepidemie 1984/85 in Osterreich.

As a consequence of the rubella epidemic in Austria in 1979, prophylaxis against this disease was intensified. In spite of this measure, however, there was an increase in cases of rubella in 1984/85. The peak incidence in both years occurred in May. In 1984, most cases were found in Vienna, Lower and Upper Austria, and Salzburg, whereas in 1985, Styria and Vienna were most affected. The clinical diagnosis of rubella was confirmed serologically in 685 patients. Of these, 151 were pregnant women and 104 were women of childbearing age who gave no information as to pregnancy. Due to the virtually complete recognition of cases, which usually led to therapeutic abortion, only 7 cases of intrauterine rubella infection were recorded. In spite of the vaccination of all prepuberal schoolgirls, of women at occupational risk, and of seronegative women after delivery, 10% of women of childbearing age still have no reliable immunity (HI titre 1:32) to rubella. This situation can be improved only by testing women at around 20 years of age for immunity (irrespective of their vaccination status) and vaccinating them if necessary.

Surveillance of viral respiratory tract infections over a one year period in mainly hospitalized Austrian infants and children by a rapid enzyme-linked immunosorbent assay diagnosis.

In order to obtain more information on viral respiratory tract infections in Austrian infants and children, nasopharyngeal secretions from 1432 infants and children, collected from October 1984 to October 1985, were screened for the presence of respiratory syncytial virus (RSV), adenoviruses, parainfluenza virus type 1, 2, and 3, and influenza viruses type A and B, by enzyme-linked immunosorbent assay (ELISA). The results obtained were analyzed with respect to incidence, seasonal distribution and clinical syndromes associated with the different viral pathogens investigated and also with the practicability of ELISA diagnostics over long distances. A viral etiology of acute respiratory tract infection was confirmed in 372 (26%) infants. RSV was detected in 286 (20%) of the nasal secretions and was thus the most frequently encountered agent. RSV infections occurred mainly in the winter months and were often associated with bronchitis, bronchiolitis, and pneumonia. Only sporadic infections were found with one of the other viruses investigated.

[Shortening of interval between first and second TBE vaccination in asthmatic children (author's transl)]. / Verkürzung des Intervalls bei FSME-Impfung bei asthmakranken Kindern.

37 children suffering from asthma had to be vaccinated against tick-borne encephalitis (TBE) with an interval of only 10 days between the first two vaccinations. Sufficient antibodies were detected in samples taken 14 days after the second injection. No differences were found between the results in this group of asthmatic children and in children who were vaccinated with the usual interval of 1 to 3 months elapsing between the first two injections. The asthmatic children tolerated the vaccination very well, moreover the indicence of side reactions was not different from that of the control group.

[Infections with respiratory viruses]. / Infektionen mit respiratorischen Viren.

Worldwide, acute infections of the respiratory tract are one of the main causes of disease, and the majority of these are due to viruses. As a consequence of the development of new methods, rapid laboratory diagnosis of virus-induced acute respiratory tract infections has greatly improved. An efficient immunoprophylaxis is still only available for influenza virus infections, and recent reports indicate that intravenous administration of immunoglobulins can protect against or reduce the severity of RS virus infections in children. At present, specific antiviral chemotherapy is only available for infections with influenza A virus and for RS-virus-infected children at high risk of a severe course of disease.

Validation of the modified hemagglutination inhibition assay (mHAI), a robust and sensitive serological test for analysis of influenza virus-specific immune response.

BACKGROUND: The hemagglutination inhibition assay (HAI) is universally regarded as the gold standard in influenza virus serology. Nevertheless, difficulties in titre readouts are common and interlaboratory variations are frequently reported. OBJECTIVE: We developed and validated the modified HAI to facilitate reliable, accurate and reproducible analysis of sera derived from influenza vaccination studies. STUDY DESIGN: Clinical and preclinical serum samples, NIBSC reference sera and seasonal influenza virus type A (H1N1 and H3N2) and type B antigens were employed to validate the mHAI. Moreover, pandemic virus strains (H5N1 and H1N1pdm09) were used to prove assay robustness. RESULTS: Utilisation of a 0.08% solution of stabilised human erythrocytes, assay buffer containing bovine serum albumin and microscopical plate readout are the major differences between the modified and standard HAI assay protocols. Validation experiments revealed that the mHAI is linear, specific and up to eightfold more sensitive than the standard HAI. In 95.6% of all measurements mHAI titres were precisely measured irrespective of the assay day, run or operator. Moreover, 96.4% (H1N1) or 95.2% (H3N2 and B), respectively, of all serum samples were determined within one dilution step of the nominal values for spiked samples. Finally, the mHAI results remained unaffected by variations in virus antigens, erythrocytes, reagents, laboratory location, sample storage conditions or matrix components. CONCLUSION: The modified HAI is easy to analyse, requires only a single source of erythrocytes and allows utilisation of numerous influenza virus antigens, also including virus strains which are difficult to handle by the standard HAI (e.g. H3N2, H5N1 and H1N1pdm09).