Gerard J. J. van Doornum

Newly discovered coronavirus as the primary cause of severe acute respiratory syndrome

Summary Background The worldwide outbreak of severe acute respiratory syndrome (SARS) is associated with a newly discovered coronavirus, SARS-associated coronavirus (SARSCoV). We did clinical and experimental studies to assess the role of this virus in the cause of SARS. Methods We tested clinical and postmortem samples from 436 SARS patients in six countries for infection with SARSCoV, human metapneumovirus, and other respiratory pathogens. We infected four cynomolgus macaques (Macaca fascicularis) with SARS-CoV in an attempt to replicate SARS and did necropsies on day 6 after infection. Findings SARS-CoV infection was diagnosed in 329 (75%) of 436 patients fitting the case definition of SARS; human metapneumovirus was diagnosed in 41 (12%) of 335, and other respiratory pathogens were diagnosed only sporadically. SARS-CoV was, therefore, the most likely causal agent of SARS. The four SARS-CoV-infected macaques excreted SARS-CoV from nose, mouth, and pharynx from 2 days after infection. Three of four macaques developed diffuse alveolar damage, similar to that in SARS patients, and characterised by epithelial necrosis, serosanguineous exudate, formation of hyaline membranes, type 2 pneumocyte hyperplasia, and the presence of syncytia. SARS-CoV was detected in pneumonic areas by virus isolation and RT-PCR, and was localised to alveolar epithelial cells and syncytia by immunohistochemistry and transmission electron microscopy. Interpretation Replication in SARS-CoV-infected macaques of pneumonia similar to that in human beings with SARS, combined with the high prevalence of SARS-CoV infection in SARS patients, fulfill the criteria required to prove that SARS-CoV is the primary cause of SARS. Published online July 22, 2003 http://image.thelancet.com/extras/03art6318web.pdf

Prevalence and Clinical Symptoms of Human Metapneumovirus Infection in Hospitalized Patients

During a 17-month period, we performed retrospective analyses of the prevalence of and clinical symptoms associated with human metapneumovirus (hMPV) infection, among patients in a university hospital in The Netherlands. All available nasal-aspirate, throat-swab, sputum, and bronchoalveolar-lavage samples (N=1515) were tested for hMPV RNA by reverse-transcriptase polymerase chain reaction. hMPV RNA was detected in 7% of samples from patients with respiratory tract illnesses (RTIs) and was the second-most-detected viral pathogen in these patients during the last 2 winter seasons. hMPV was detected primarily in very young children and in immunocompromised individuals. In young children, clinical symptoms associated with hMPV infection were similar to those associated with human respiratory syncytial virus (hRSV) infection, but dyspnea, feeding difficulties, and hypoxemia were reported more frequently in hRSV-infected children. Treatment with antibiotics and corticosteroids was reported more frequently in hMPV-infected children. From these data, we conclude that hMPV is an important pathogen associated with RTI.

A cluster of acute hepatitis C virus infection among men who have sex with men : results from contact tracing and public health implications

Objective:An acute hepatitis C virus (HCV) infection in an HIV-positive man who had sex with men (MSM) was notified. In the period of his seroconversion he was also diagnosed with a rectal lymphogranuloma venereum (LGV) infection, and was part of a cluster of 15 LGV cases in 2003. Our aim was to investigate HCV transmission and to search for potential spread among sexual contacts and known LGV patients. Methods:Our case series included the index, two recent contacts, and 14 LGV cases. They were interviewed about parenteral exposure for HCV, history of sexually transmitted diseases(STDs), sexual behaviour and drug use. Laboratory investigations included anti-HCV antibodies, HCV-polymerase chain reaction, and HCV genotyping. Results:Seven out of 17 MSM recently seroconverted for HCV (41%). Three genotypes were found. Parenteral risk factors were excluded. Six out of seven had LGV proctitis coinciding with HCV seroconversion, six (86%) were HIV infected. Unprotected anal contact was practised by both HCV uninfected and infected cases. Unprotected active and passive fisting was reported by all seven HCV infected men, compared with two of nine uninfected men (P = 0.003). Non-intravenous drug use during sexual activities was common among all MSM. Numerous, often anonymous, sexual contacts in various European countries were reported. Conclusions:A cluster of acute HCV infection is reported among mostly HIV-positive MSM, with multiple partners throughout Europe. Sexual techniques potentially leading to mucosal damage (fisting), concomitant STDs such as LGV and drug use seem facilitating factors for spread. Extensive case finding and partner tracing is advocated as well as targeted prevention messages.

Modified Vaccinia Virus Ankara Protects Macaques against Respiratory Challenge with Monkeypox Virus

ABSTRACT The use of classical smallpox vaccines based on vaccinia virus (VV) is associated with severe complications in both naïve and immune individuals. Modified vaccinia virus Ankara (MVA), a highly attenuated replication-deficient strain of VV, has been proven to be safe in humans and immunocompromised animals, and its efficacy against smallpox is currently being addressed. Here we directly compare the efficacies of MVA alone and in combination with classical VV-based vaccines in a cynomolgus macaque monkeypox model. The MVA-based smallpox vaccine protected macaques against a lethal respiratory challenge with monkeypox virus and is therefore an important candidate for the protection of humans against smallpox.

Human papillomavirus infection in men who have sex with men participating in a Dutch gay-cohort study

Background To develop strategies for prevention and early treatment of human papillomavirus (HPV) anal and penile cancer, a better understanding of related sexual behavior risk factors is needed. Goal The goal of this study was to establish the prevalence of anal and coronal sulcus HPV in a group of men who have sex with men participating in a Dutch gay-cohort study, to identify risk factors associated with HPV infection in this group, and to investigate the presence of identical HPV types in couples with stable relationships. Study Design A cross-sectional study of 241 HIV-negative and 17 HIV-positive men who have sex with men visiting the sexually transmitted disease clinic of the Erasmus MC for a regular and scheduled examination. Participants underwent a routine venereological examination including HIV serologic analysis, and swabs were taken from the coronal sulcus and anus for HPV DNA testing. All subjects were asked to complete a questionnaire on sexual risk behavior. Results HPV DNA was detected at the coronal sulcus in 23.5% of the HIV-positive men and in 15.8% of the HIV-negative men (P = 0.492). In anal specimens, HPV DNA was detected in 64.7% of the HIV-positive men and 32.8% of the HIV-negative men (P = 0.015). High-risk HPV types (P = 0.007) and 2 or more different HPV genotypes (P = 0.006) were seen more often in anal specimens of HIV-positive persons than in specimens of HIV-negative persons. A factor possibly associated with the presence of anal HPV infection was a concomitant anal infection with Chlamydia trachomatis, gonococci, or herpes simplex virus (P = 0.059). In only 16.7% of HPV-positive steady couples, both companions showed the presence of one or more identical HPV genotypes. Conclusion In this study, anal HPV DNA was detected more often than HPV DNA at the coronal sulcus. HIV positivity was associated with a higher prevalence of high-risk, but not with low-risk HPV types, at the anus. No association was found between HIV positivity and presence of high-risk HPV at the coronal sulcus. No sexual behavioral determinants for the presence of HPV could be identified. Concomitant anal infection with C trachomatis, gonococci, or herpes simplex virus may be associated with HPV infection. In the majority of steady couples, partners were infected with different HPV types.

Emerging group-A rotavirus and a nosocomial outbreak of diarrhoea

A P[6]G9 group-A rotavirus caused a protracted hospital outbreak of neonatal diarrhoea in The Netherlands. The outbreak lasted 5 months with 52 cases and an average attack rate of 40%, 46 cases were in an incubator section for neonates under 1 month of age. Rotavirus P161G9 was detected by RT-PCR in stool samples from the 31 cases tested. Emergence of this genotype in Europe may have implications for neonates lacking protective maternal antibodies and for the development of rotavirus vaccines.

Fatal human rabies due to duvenhage virus from a bat in Kenya: failure of treatment with coma-induction, ketamine, and antiviral drugs.

11Department of Infectious Diseases, Tropical Medicine and AIDS, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands, 2Department ofNeurology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands, 3Department of Intensive Care, Academic Medical Center, University ofAmsterdam, Amsterdam, The Netherlands, 4Department of Clinical Virology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands,5Department of Virology, Erasmus Medical Centre, Rotterdam, The Netherlands, 6Department of Radiology, Academic Medical Center, University of Amsterdam,Amsterdam, The Netherlands, 7Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands, 8Centre of Expertise forRabies, Canadian Food Inspection Agency, Ottawa Laboratory (Fallowfield), Ottawa, Ontario, Canada

Monitoring cytomegalovirus IE-1 and pp65-specific CD4+ and CD8+ T-cell responses after allogeneic stem cell transplantation may identify patients at risk for recurrent CMV reactivations.

We studied the recovery of CMV‐specific CD4+ and CD8+ T‐cell immunity in 52 recipients of allogeneic stem cell transplantation (SCT). The proportions of IFN‐γ‐producing CD4+ and CD8+ T cells upon in vitro activation using peptide pools representing the CMV pp65 and IE‐1 proteins were assessed at multiple time points post SCT, and correlated with the occurrence of CMV reactivation. In a retrospective analysis, recurrent CMV reactivations occurred in 9 patients and were associated with low pp65‐specific CD4+ T‐cell and low IE‐1‐specific CD8+ T‐cell reactivities, whereas patients without detectable CMV reactivation (n = 30) or a single reactivation (n = 13) showed a better recovery of these immune responses. CD4+ T‐cell responses to IE‐1 were infrequent in most patients, whereas CD8+ T‐cell responses to pp65 occurred frequently, but did not correlate with protection against (recurrent) reactivation. Prospectively, CMV‐specific T‐cell responses could be studied prior to 14 reactivation episodes in 8 patients. CD4+ T‐cell responses to IE‐1 and pp65 were positive in only 1 and 2 episodes, respectively. CD8+ T‐cell responses against IE‐1 were positive in 4, but against pp65 in 12 episodes, again showing that CD8+ T‐cell reactivity against pp65 did not prevent CMV reactivation. Thus, monitoring of particular CMV‐specific CD4+ and CD8+ T‐cell responses after allogeneic SCT may identify patients at risk for recurrent CMV reactivations.

A Prospective Open Study of the Efficacy of High-Dose Recombinant Hepatitis B Rechallenge Vaccination in HIV-Infected Patients

Double-dose hepatitis B virus revaccination of human immunodeficiency virus (HIV)-infected patients proved to be effective in 50.7% of 144 patients who had previously failed to respond to standard doses. In the multivariate analysis, female patients were found to have a significantly better response (P= .03). The effect of age on the response depended on the viral load at the time of revaccination. For patients with a detectable HIV RNA load, the effect of age was stronger (odds ratio [OR], 0.34 per 10 years older [95% confidence interval {CI}, 0.16-0.72]; P= .005) than for patients with an undetectable HIV RNA load (OR, 0.74 per 10 years older [95% CI, 0.50-1.09]; P= .12).

An Outbreak of Diarrhea in a Neonatal Medium Care Unit Caused by a Novel Strain of Rotavirus: Investigation Using Both Epidemiologic and Microbiological Methods

OBJECTIVE In December 1999, an outbreak of diarrhea was reported in a general hospital neonatal medium care unit (NMCU) caused by a novel strain of rotavirus with genotype P[6], G9. An investigation was conducted to determine risk factors for illness among neonates. DESIGN Rotavirus diagnosis was by latex agglutination and typing by reverse transcriptase polymerase chain reaction. A case-control study was performed using data collected from medical records on exposures in a 3-day period before illness (cases) or a random 3-day period (controls). Environmental swabs were tested for rotavirus. Antenatal blood samples from mothers and blood samples provided by hospital staff were analyzed for rotavirus antibodies. RESULTS Fifty-six cases of rotaviral illness were confirmed by latex agglutination. Forty-seven of these were among 118 neonates exposed to the NMCU (attack rate, 40%). There was a 4-week period with no clinical cases in the course of the outbreak. Increased frequency (> or = 15 times in 3 days) of ungloved nasogastric feeding was a significant risk factor (adjusted odds ratio, 8.79), controlling for birth weight and gestational age. Environmental sampling showed persistence of the virus on ward surfaces despite cleaning. None of 24 NMCU staff members had high levels of antibodies against P[6], G9. Three (8%) of 38 mothers had high antibody levels; 2 had infants who became ill. The outbreak ended with a 7-day ward closure, disinfection, and introduction of gloved nasogastric feeding. CONCLUSIONS Case-control studies can be successful in identiffying risk factors for nosocomial outbreaks of diarrhea. High levels of rotavirus antibodies in mothers may not protect infants. The environment may be the most important reservoir of rotavirus during outbreaks.