Infections du système nerveux central à entérovirus: 223 cas vus à un h?pital pédiatrique entre 1973 et 1981.

Between 1973 and 1981, 223 patients were seen at hôpital Sainte-Justine in Montreal for enteroviral infection of the nervous system. In 161 the cause was documented by isolation of an enterovirus from the cerebrospinal fluid (CSF). The viruses most frequently isolated were echovirus 11 (36 isolates), echovirus 30 (24), coxsackievirus B2 (23), coxsackievirus B3 (19), echovirus 6 (18), coxsackievirus B5 (16), coxsackievirus A9 (15), echovirus 9 (13), echovirus 7 (12) and coxsackievirus B1 (11). Aseptic meningitis was diagnosed in 200 cases and encephalitis in 12. The remaining 11 patients presented with the features of septicemia or with convulsions. In 33 patients an enterovirus was isolated from the CSF in the absence of pleocytosis. Polymorphonuclear cell predominance was noted in the initial CSF sample in 95 cases; it was persistent in 11. There were five mixed infections; in three cases two viruses were isolated from the same CSF sample. Two patients died: one, a child with hypogammaglobulinemia, had fatal polioencephalitis; the other, a 6-month-old infant brought to the emergency room in unexplained cardiopulmonary arrest, had echovirus 6 meningitis. Of the 172 patients admitted to hospital 96 received parenteral antibiotic therapy. The impact of enteroviral infections of the central nervous system on hospital resources could be substantially reduced if a rapid, sensitive and specific laboratory method of diagnosing these infections were available.     

Enteroviral aseptic meningitis in Japan, 1981-1991. A report of the National Epidemiological Surveillance of Infectious Agents in Japan.

In Japan, aseptic meningitis cases due to enterovirus infections increase every summer in various degrees with an incidence peak usually in July. During the past 11 years from 1981 through 1991, a total of 8,595 enterovirus isolations from aseptic meningitis cases were reported from 54 participating laboratories. Eight enterovirus types caused large epidemics; more than 100 isolations of each type from aseptic meningitis cases were reported for every epidemic year of the respective type. They were coxsackievirus (C) types B3 and B5, echovirus (E) types 4, 6, 7, 9, 18 and 30. Among these, the highest meningitis-associating frequency was reported for E30, representing 82.6% of the total isolations reported for the type during this period, followed by E4, 71.1%. The frequencies of E9, E7, E6 and CB5 were in a range from 54.5% to 44.4%, while that of E18 was 37.7% and that of CB3 21.0%. During the epidemics, enterovirus-associated meningitis was most frequently reported among children of 4-7 years of age. High frequencies were also shown in infants less than 1-year of age in some types. A total of 4,240 enteroviruses were isolated from cerebrospinal fluid of aseptic meningitis cases, representing 49.3% of the cases with enterovirus isolation.     

Characterization of an echovirus type 30 variant isolated from patients with aseptic meningitis

An outbreak of echovirus type 30 infection associated with aseptic meningitis occurred among newborn babies in a hospital neonatal room at Fukui City in 1983. The isolated virus was identified as an antigenic variant of echovirus type 30 by cross-neutralization tests with antisera against the prototype Bastianni strain and the present isolate. Western blot analysis demonstrated that the antigenic determinants responsible for virus neutralization, some of which were type specific and others strain specific, were present on the capsid protein VP1. The current strain was more thermoresistant than the prototype, suggesting some alterations in virus structural proteins.     

Molecular epidemiology of echoviruses 11 and 13, based on an environmental surveillance conducted in Toyama Prefecture, 2002-2003

Nineteen echovirus 11 (E11) and 12 E13 isolates were isolated from three rivers in Toyama Prefecture, Japan, during an environmental surveillance conducted from April 2002 to March 2003. The nucleotide sequences of E13 isolates were closely related to those from patients with aseptic meningitis, with less than 1.3% divergence in the VP1 region of the viral capsid gene, and belonged to the same clade responsible for a worldwide outbreak that started in 2000. In contrast, E11 isolates were clustered into three genomic groups and were not closely related to echovirus strains isolated from patients. These results suggest that the combination of both virus isolation from environmental sources and phylogenetic analysis could be complementary assessment approaches to trace prevalent and minor circulating enteroviruses in the human population.     

Molecular Epidemiology of Echoviruses 11 and 13, Based on an Environmental Surveillance Conducted in Toyama Prefecture, 2002-2003

Nineteen echovirus 11 (E11) and 12 E13 isolates were isolated from three rivers in Toyama Prefecture, Japan, during an environmental surveillance conducted from April 2002 to March 2003. The nucleotide sequences of E13 isolates were closely related to those from patients with aseptic meningitis, with less than 1.3% divergence in the VP1 region of the viral capsid gene, and belonged to the same clade responsible for a worldwide outbreak that started in 2000. In contrast, E11 isolates were clustered into three genomic groups and were not closely related to echovirus strains isolated from patients. These results suggest that the combination of both virus isolation from environmental sources and phylogenetic analysis could be complementary assessment approaches to trace prevalent and minor circulating enteroviruses in the human population.     

Mumps and Enteroviral Meningitis in Toronto, 1966

Of 52 children admitted to hospital for apparently typical mumps meningitis in 1966, 50 had their cerebrospinal fluid (CSF) examined. In only 17 was the mumps virus isolated from the CSF. Mumps antihemagglutinin conversions or increments were detected in 32 subjects including 10 whose CSF yielded virus. Antibody conversions were found in 16 patients and fourfold increments in another nine whose serum pairs were collected only one to four days apart. Initial sera from 20 patients were obtained three days or less after the onset of meningitis. Antibody increments were frequently noted about one day after defervescence and clinical improvement. Interferon was detected in CSF from two of eight patients, both of whom yielded virus. Enteroviruses were isolated from CSF and/or feces in seven of 15 cases of aseptic meningitis which occurred between July and October. Six patients including three virus excretors showed enteroviral neutralizing antibody increments during convalescence. The dominant enteroviral serotype was coxsackievirus A9.     

Outbreak of aseptic meningitis by echo 4: prevalence of clinical cases among adults

Twenty five cases of meningitis occurred in urban areas surrounding a city (Modena) in Northern Italy, in the period May-July 1999. When the patients were admitted to the Infectious Diseases Division of the University of Modena and Reggio Emilia Hospital and studied by virological and serological methods, the meningitis proved to have an enteroviral origin and enterovirus ECHO 4 type was responsible for all cases of illness. An epidemiological characteristic of the enteroviral meninigitis outbreak was the adult age in 23 out of the 25 patients (mean age 24.50 +/- 7.84 years). The monthly distribution of the aseptic meningitis cases was the following: five cases occurred in May, 13 in June and seven in July. The origin of the spread of the virus infection and the reason for its sudden end remained unknown. The unusual drop in temperature which occurred in the geographic area involved in the aseptic meningitis outbreak at the beginning of August could have interfered with the slowdown in virus circulation.     

Use of monoclonal antibody blends in the identification of enteroviral aseptic meningitis

Monoclonal antibody pools directed against group B coxsackievirus and echovirus antigens (Chemicon, Temecula, California) were evaluated as tools in the identification of enteroviral aseptic meningitis. Cerebrospinal fluid (CSF) and serial dilutions of stock coxsackievirus were inoculated into tube and shell vial Rhesus monkey kidney (RhMk) cell cultures. Positive cellular fluorescence was observed only in conjunction with cytopathic effect (CPE). The time from inoculation to CPE was similar with both tube and shell vial cultures.Direct CSF testing failed to reliably identify positive specimens as fluorescent debris, and a lack of available cells hindered results.Viral components of each antibody blend demonstrated positive cellular fluorescence when appropriately stained. False-positive fluorescence was not observed when cells, infected with other CSF viruses, were stained with these reagents.The findings suggest a role for these reagents (available both as blends and type-specific reagents) in the culture confirmation and identification of many common enteroviral serotypes associated with aseptic meningitis.     

Elucidation of Echovirus 30's Origin and Transmission during the 2012 Aseptic Meningitis Outbreak in Guangdong,China, through Continuing Environmental Surveillance

An aseptic meningitis outbreak occurred in Luoding City of Guangdong, China, in 2012, and echovirus type 30 (ECHO30) was identified as the major causative pathogen. Environmental surveillance indicated that ECHO30 was detected in the sewage of a neighboring city, Guangzhou, from 2010 to 2012 and also in Luoding City sewage samples (6/43, 14%) collected after the outbreak. In order to track the potential origin of the outbreak viral strains, we sequenced the VP1 genes of 29 viral strains from clinical patients and environmental samples. Sequence alignments and phylogenetic analyses based on VP1 gene sequences revealed that virus strains isolated from the sewage of Guangzhou and Luoding cities matched well the clinical strains from the outbreak, with high nucleotide sequence similarity (98.5% to 100%) and similar cluster distribution. Five ECHO30 clinical strains were clustered with the Guangdong environmental strains but diverged from strains from other regions, suggesting that this subcluster of viruses most likely originated from the circulating virus in Guangdong rather than having been more recently imported from other regions. These findings underscore the importance of long-term, continuous environmental surveillance and genetic analysis to monitor circulating enteroviruses.     

An Outbreak of Aseptic Meningitis due to Echo 25 Virus

In a widespread outbreak of aseptic meningitis during the summer and early fall of 1961 in Ontario, Echo type 25 virus was found to be the predominant virus type isolated from affected patients. Recovery of Echo 25 virus in several cases from the CSF of patients strongly suggests an etiological relationship between this virus type and the illness. The causative role of the virus was confirmed by demonstration of significant immunological response in patients found to be infected with the virus. The presence of antibodies to Echo 25 virus in about 70% of meningitis patients, from whom no virus was isolated, indicates that the infection in the population of the affected areas was much more prevalent than the number of successful virus isolations would suggest. It is recommended that Echo 25 virus should be added to the series of Echo virus types capable of causing outbreaks of aseptic meningitis.     

Assessment of an enterovirus sewage surveillance system by comparison of clinical isolates with sewage isolates from milwaukee, wisconsin, collected august 1994 to december 2002

The quantity and serotypes of enteroviruses (EVs) in the influent of a local sewage treatment plant were compared to local clinical EV cases to determine if testing of sewage is adequate for an EV surveillance system. The study was carried out from August 1994 to December 2002. Monthly influent specimens were processed by organic flocculation, and dilutions of concentrate were inoculated onto a number of different cell types for virus isolation. EVs were detected in 88 of 100 monthly influent samples. Sewage EV titers were calculated by using software provided by the U.S. Environmental Protection Agency for most-probable-number determination. All 1,068 sewage EV isolates were further grouped (echovirus, coxsackievirus B, coxsackievirus A, or poliovirus) by cell culture host range analysis (growth pattern of isolates on passage to seven cell lines), and 39.0% of the 1,022 EV isolates categorized as non-poliovirus EVs were specifically serotyped. For clinical cases, primary virus isolation tests were performed on specimens submitted by local hospitals and EV isolates submitted by hospitals were serotyped. Clinical EVs were documented for 81 of the 100 months studied. In all, 694 EV isolates from clinical cases were serotyped. Annually, between 4 and 11 different serotypes of non-poliovirus EVs were identified in sewage and from 9 to 19 different non-poliovirus EV serotypes were identified from clinical specimens. Usually, the most commonly detected sewage EV serotypes were similar to the most commonly detected clinical serotypes; e.g., for 1997, echovirus 6 accounted for 53.1% of the typed sewage isolates and 39.4% of the clinical infections, while in 1998, echovirus 30 accounted for 50.0 and 46.1%, respectively. In 1999, 60.3% of the EVs from clinical cases and 79.7% of the sewage isolates were echovirus 11; in 2000, 33.3% of the EVs from clinical cases and 40.7% of the sewage isolates were coxsackievirus B5; and in 2001, 44.1% of the EVs from clinical cases and 36.2% of the sewage isolates were echovirus 13. Annual peaks of both sewage EV titers and clinical cases occurred in late summer or early fall. In some years, early spring sewage EVs portended some of the EVs that would predominate clinically during the following summer.     

Assessment of an Enterovirus Sewage Surveillance System by Comparison of Clinical Isolates with Sewage Isolates from Milwaukee, Wisconsin, Collected August 1994 to December 2002

The quantity and serotypes of enteroviruses (EVs) in the influent of a local sewage treatment plant were compared to local clinical EV cases to determine if testing of sewage is adequate for an EV surveillance system. The study was carried out from August 1994 to December 2002. Monthly influent specimens were processed by organic flocculation, and dilutions of concentrate were inoculated onto a number of different cell types for virus isolation. EVs were detected in 88 of 100 monthly influent samples. Sewage EV titers were calculated by using software provided by the U.S. Environmental Protection Agency for most-probable-number determination. All 1,068 sewage EV isolates were further grouped (echovirus, coxsackievirus B, coxsackievirus A, or poliovirus) by cell culture host range analysis (growth pattern of isolates on passage to seven cell lines), and 39.0% of the 1,022 EV isolates categorized as non-poliovirus EVs were specifically serotyped. For clinical cases, primary virus isolation tests were performed on specimens submitted by local hospitals and EV isolates submitted by hospitals were serotyped. Clinical EVs were documented for 81 of the 100 months studied. In all, 694 EV isolates from clinical cases were serotyped. Annually, between 4 and 11 different serotypes of non-poliovirus EVs were identified in sewage and from 9 to 19 different non-poliovirus EV serotypes were identified from clinical specimens. Usually, the most commonly detected sewage EV serotypes were similar to the most commonly detected clinical serotypes; e.g., for 1997, echovirus 6 accounted for 53.1% of the typed sewage isolates and 39.4% of the clinical infections, while in 1998, echovirus 30 accounted for 50.0 and 46.1%, respectively. In 1999, 60.3% of the EVs from clinical cases and 79.7% of the sewage isolates were echovirus 11; in 2000, 33.3% of the EVs from clinical cases and 40.7% of the sewage isolates were coxsackievirus B5; and in 2001, 44.1% of the EVs from clinical cases and 36.2% of the sewage isolates were echovirus 13. Annual peaks of both sewage EV titers and clinical cases occurred in late summer or early fall. In some years, early spring sewage EVs portended some of the EVs that would predominate clinically during the following summer.     

Isolation and Characterization of Mumps Virus Strains in a Mumps Outbreak with a High Incidence of Aseptic Meningitis

In 1993, mumps with a high incidence of aseptic meningitis became prevalent in Akita prefecture, Japan. Three mumps virus isolates obtained from the nonvaccine-associated cases lacked the BamHI restriction cleavage site of the P gene, like the Urabe strain (Yamada, A. et al, Vaccine 8: 553-557). However, four additional nucleotide substitutions were found in the determined region of 157 bp. Fourteen of 19 cases from which mumps virus showing the Urabe-like RFLP profile was detected were complicated with symptomatic meningitis, whereas there were only four cases of meningitis among 23 individuals infected with the wild type showing no Urabe-like RFLP profile (non-“Urabe-like” wild-type). The incidence of meningitis was over 70% among patients infected with the “Urabe-like” wild-type virus. The “Urabe-like” wild-type disappeared after February 1994 in the epidemic area and was replaced by the non-“Urabe-like” wild-type. Patients infected with the “Urabe-like” wild-type lived in a closed colony, in which there were two instances of transmission between siblings. Thus this outbreak was transient and narrowly localized.     

Molecular detection and characterization of human enteroviruses in Korean surface water

In this study, the genetic epidemiology of enteroviruses (EVs) in Korean surface water was evaluated by conducting phylogenetic analyses of the nucleotide sequences of the 5' non-coding region (5' NCR), which was determined by RT-PCR analysis of total culturable virus assay-positive samples. The results showed that the nucleotide sequences of the EVs could be classified into 4 genetic clusters, and that the predominant presence of Korea EVs were very similar to echoviruses type 30. Interestingly, two nucleotide sequences were very similar to those of coxsackievirus type B1 isolated from aseptic meningitis patients in Seoul, Korea, implying the possibility of a common source for the viruses circulated in water systems and humans. In addition, 3 nucleotide sequences clustered strongly with the nucleotide sequences from China or Japan, and one fell into the same cluster as echovirus type 11 from Taiwan, which suggests that EVs in Asia may have evolved in a region-specific manner. Taken together, the results of this study revealed that EVs from Korea surface waters could be genetically classified as coxsackieviruses or echoviruses, and that they evolved in Asia in a region-specific manner.     

A clustering outbreak of hand, foot, and mouth disease caused by Coxsackie virus A10

A clustering outbreak of hand, foot, and mouth disease (HFMD) occurred from July, 1981 to January, 1982 in Matsue City and Gotsu City, Shimane Prefecture. Thirty-seven patients with clinical HFMD were virologically and serologically examined, and Coxsackie virus A10 (CA10) was isolated in 18 patients from vesicles (7/16), throat-swabs (9/31) and feces (6/7). During the period, no CA16 or enterovirus 71 were isolated from HFMD patients or from other diseases such as pharyngitis, febrile diseases, and aseptic meningitis. Serological diagnosis was performed employing an African green monkey kidney cell (AG-1)-adapted CA10 which demonstrated cytopathogenic effects on the cells. Paired sera from seven patients including three cases in which isolation failed showed a significant increase of neutralizing antibody titer against CA10. Finally, an etiological diagnosis was made in 21 out of 37 patients with clinical HFMD. This is the first report of a clustering outbreak of HFMD caused by CA10 in Japan.     

Enteroviral and Mumps Meningitis in Toronto, 1963

Virological investigations of 115 children with the aseptic meningitis syndrome during 1963 resulted in the isolation of enteroviruses from cerebrospinal fluid (CSF) and/or feces of 21 of 48 children who had no association with mumps. For the third successive year, Echo 9 was the dominant enterovirus in cases of aseptic meningitis in Toronto children, but no rashes were associated with Echo 9 meningitis during 1963, in contradistinction to previous years. Mumps virus was isolated from CSF of 25 patients by inoculation of rhesus monkey kidney cultures, and rising or elevated mumps antihemagglutinin titres in paired sera from a further 33 cases provided laboratory evidence of infection with mumps virus in 58 of 67 patients with mumps meningoencephalitis. No enlargement of salivary glands was noted in 20 laboratory-proved cases of mumps meningoencephalitis. Enteroviral meningitis occurred principally during summer, but the peak of mumps meningoencephalitis occurred during late winter.     

Rash Associated with Coxsackie A9 Infection

Coxsackie A9 virus was identified by the authors during the fall of 1965 in Montreal in six children with fever and exanthem. Three of the six children were siblings. The exanthem was centrally distributed as described by Lerner et al. and consisted of discrete maculopapules 3 to 4 mm. in diameter. The viral agent was recovered and identified in tissue culture in five cases, while in the sixth Coxsackie type-A lesions were produced in suckling mice. Serological confirmation was obtained in two patients from whom sera were available. In contrast, no exanthem was observed in three older patients with a diagnosis of aseptic meningitis associated with Coxsackie A9 virus. In only one of 16 patients with Coxsackie B virus infection was an exanthem observed during the same period.The true incidence of Coxsackie A9-associated exanthems is difficult to determine because of the benign nature of the disease.     

Environmental Surveillance of Human Enteroviruses in Shandong Province,China, 2008 to 2012: Serotypes,Temporal Fluctuation,and Molecular Epidemiology

Environmental surveillance is an effective approach in investigating the circulation of polioviruses (PVs) and other human enteroviruses (EVs) in the population. The present report describes the results of environmental surveillance conducted in Shandong Province, China, from 2008 to 2012. A total of 129 sewage samples were collected, and 168 PVs and 1,007 nonpolio enteroviruses (NPEVs) were isolated. VP1 sequencing and typing were performed on all isolates. All PV strains were Sabin-like, with the numbers of VP1 substitutions ranging from 0 to 7. The NPEVs belonged to 19 serotypes, and echovirus 6 (E6), E11, coxsackievirus B3 (CVB3), E3, E12, and E7 were the six main serotypes, which accounted for 18.3%, 14.8%, 14.5%, 12.9%, 9.0%, and 5.7% of NPEVs isolated, respectively. Typical summer-fall peaks of NPEV were observed in the monthly distribution of isolation, and an epidemic pattern of annual circulation was revealed for the common serotypes. Phylogenetic analysis was performed on environmental CVB3 and E3 strains with global reference strains and local strains from aseptic meningitis patients. Shandong strains formed distinct clusters, and a close relationship was observed between local environmental and clinical strains. As an EV-specific case surveillance system is absent in China and many other countries, continuous environmental surveillance should be encouraged to investigate the temporal circulation and phylogeny of EVs in the population.     

Viral Infections of Toronto Children During 1965: I. Enteroviral Disease

Enteroviruses were isolated from feces and/or cerebrospinal fluid of 29 of 43 Toronto children who contracted aseptic meningitis, pleurodynia, abdominal pain or febrile upsets between June and October, 1965. Coxsackie A9 virus was the dominant agent in aseptic meningitis and Coxsackie B1 virus in pleurodynia and other syndromes. Sero-logical evidence of recent Coxsackie B1 and Echo 6 infection was obtained in two additional patients with aseptic meningitis who did not yield virus, and elevated Coxsackie B1 antibody titres were found in one patient with pericarditis. A newborn infant died with myocarditis due to Coxsackie B1 virus following infection of the mother during the immediate antenatal period. Paired sera collected only two to four days apart from patients with enteroviral syndromes or mumps meningoencephalitis frequently showed four-fold or greater increases of antibody levels.     

Primary Culture of Human Embryonic Kidney Cells as the Medium for Isolation of ECHO 6 Virus

A large epidemic of aseptic meningitis due to ECHO 6 virus swept over Japan in the summer months of 1965. In our studies on 89 cases of aseptic meningitis in the epidemic, primary culture of human embryonic kidney cells was shown to provide a highly sensitive host system for isolation of ECHO 6 virus from clinical materials. Virus was recovered from cerebrospinal fluid in 72% of 89 cases, from throat swabs in 64% of 36 cases, and from rectal swabs in 59% of 37 cases. Most significant is the finding that the rate of virus isolation from cerebrospinal fluid in this host system was considerably higher as compared with that obtained by other investigators in other host systems such as primary monkey kidney cells or human amniotic cells (primary or EL). This finding should be emphasized particularly because isolation of a virus from cerebrospinal fluid, in contrast to throat secretions or feces, is of much greater importance in establishing the etiologic relationship to the disease in the diagnosis of aseptic meningitis. The neutralization test was shown to be efficient in detecting ECHO 6 virus infection; acute serums should be taken preferably by the 4th day of illness and convalescent serums in the second week. Epidemiologic findings, such as the predominance of male patients and occurrence of the epidemic in summer months, generally coincide with the previous reports. However, our cases were in much younger age groups in contrast with the previous reports; 90% of our 89 patients were 6 years of age or younger, and 15 children or 17% were less than 1 year of age, including 3 less than 6 months of age. The clinical observations on our cases confirmed the previous reports.