Echovirus 6, Human ; genetics ; isolation & purification ; Enterovirus Infections ; virology ; Genes, Viral ; Humans ; Sequence Analysis

BACKGROUND: Aseptic meningitis is a common illness of children. It seems that viruses are the usual etiologic agents. The distribution of these agents mainly depends on the isolated time and region area. This study was performed to isolate the causative viruses from patients with aseptic meningitis in Gwangju area during recent one year. METHODS:A total of 130 patients with aseptic meningitis were evaluated. Stool and/or cerebrospinal fluid specimens from patients were inoculated into rhabdomyosarcoma (RD), HEp2 and Vero cell lines. The virus propagation was examined by the presence of cytopathic effects. Neutralizing tests using enterovirus serum pool were done on each viral isolates. RESULTS: The isolation rate of enterovirus was 24.6% (32/130). The enterovirus isolates were obtained mostly from stool specimens (29/32). Twenty-two isolates were identified by neutralizing test. Ten isolates disclosed 'untyping' by neutralizing test. The distribution of isolates was coxsackievirus group B2 (11 stains, 34.4%), echovirus 30 (4 strains, 12.5%), echovirus 6 (3 strains, 9.4%), echovirus 9, 11, 25 and coxsackievirus group A16 (1 strain, respectively). These strains were predominantly isolated during summer season (June to July). CONCLUSIONS: The causative viruses from patients with aseptic meningitis in Gwangju area during recent 1 year were coxsackievirus group B2, echovirus 30, 6, 9, 11, 25 and coxsackievirus group A16 which were mostly isolated from stool specimens in summer season.
Cerebrospinal Fluid ; Child ; Coloring Agents ; Echovirus 6, Human ; Echovirus 9 ; Enterovirus ; Enterovirus B, Human ; Gwangju* ; Humans ; Meningitis, Aseptic* ; Rhabdomyosarcoma ; Seasons ; Vero Cells

PURPOSE: There was an outbreak of aseptic meningitis in Busan and Kyoungsangnamdo area from April to July, 2002. In this study, we reported the clinical manifestations, laboratory data and causative viruses. METHODS: 184 children with aseptic meningitis who had been admitted to Department of Pediatrics, Inje Paik Hospital in Kaegum and Dongrae between April and July 2002 were assessed. Virus isolation and serotype identification were performed by cell culture and reverse transcription-polymerase chain reaction(RT-PCR) of cerebrospinal fluid. RESULTS: The peak incidence was noted in May. Male-to-female ratio was 2:1. The age ranged from 2 years to 16 years. Mean age was 8.7+/-7.1 years. Clinical manifestations were fever(95.1%), headache(91.8%) and vomiting(89.7%). Mean duration of fever was 2.6 days. The numbers of peripheral blood leukocytes were 1,400-20,000/mm3 and mean value was 9,829/mm3. On cerebrospinal fluid examinations, leukocyte count was in range of 10-2,000(mean 301)/mm3, protein level in range of 10-196(mean 36.7)mg/dL and glucose level from 17 to 155(mean 58.3)mg/dL. Viruses were isolated 13(8.4%) out of 154 patients and serotypes were 6 cases of echovirus 6, 3 cases of echovirus 9, 1 case of echovirus 13, 2 cases of echovirus 25 and 1 case of echovirus 30. CONCLUSION: Aseptic meningitis was prevalent in Busan and Kyoungsangnamdo area from April to July, 2002. We thought that causative viruses were echovirus 6, 9, 13, 25, 30.
Busan* ; Cell Culture Techniques ; Cerebrospinal Fluid ; Child ; Echovirus 6, Human ; Echovirus 9 ; Enterovirus B, Human ; Fever ; Glucose ; Gyeongsangnam-do* ; Humans ; Incidence ; Leukocyte Count ; Leukocytes ; Meningitis, Aseptic* ; Pediatrics

This study aimed to analyze the etiology of the encephalitis outbreak in Longyan, Fujian Province, China in 2010, in order to provide valuable information for this prevention and control of this disease. Pathogens were confirmed from cerebrospinal fluid samples with fluorescent RT-PCR, virus isolation (RD cells), and neutralization tests. Then, the VP1 fragments or whole genome nucleotide sequences were determined for four virus strains using PCR. Homology was assessed using the MegAlign software, and a phylogenetic evolutionary tree was drawn using Mega 4.0 software. The results confirmed that the etiology of the outbreak was the ECHO6 intestinal virus, and the nucleotide sequence of the VP1 segment indicated that the C2 subtype was responsible. The genome sequence consisted of 7407 nucleotides, and resembled the genome of other ECHO and CoxB viruses with homology levels of 78.5%-87.3%. The encephalitis outbreak in Longyan in 2010 was caused by the ECHO6 C2 subtype intestinal virus, and its complete genome sequence length is similar to the standard strain (U16283) with a sequence homology of 80.4%.
Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Echovirus 6, Human ; classification ; genetics ; isolation & purification ; Echovirus Infections ; epidemiology ; virology ; Encephalitis ; epidemiology ; virology ; Female ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny

OBJECTIVETo understand the epidemiological, clinical and etiological characteristics of an Echovirus type 6 meningitis outbreak in Jinzhai county, Liu'an city in Anhui, and to find out the proper way in controlling the aseptic meningitis outbreak.

METHODSA surveillance system for aseptic meningitis was established in Jinzhai to confirm the case definition. Stool or cerebrospinal fluid (CSF) specimens from some cases were collected for entero-viruses isolation and identification. Case-control study was conducted. The case group involved patients while the controls would include: patients' classmate with same gender and the age difference was not over one year. Neutralization antibody in serum specimens were collected and tested in cases and in healthy people.

RESULTS105 cases were distributed in 17 of the 30 towns in Jinzhai county while 41.0% of the cases were in Banzhuyuan town with an incidence rate of 203/10(5). Cases were clustered by school and classroom with age ranging from 3 to 15 years old and the highest as 10.9/10(5) in the 6 to 10 group. The incidence in males was 24.2/10(5) compared to 8.4/10(5) in females. The main clinic characteristics of cases were: fever, headache and vomiting. Echovirus type 6 from 25 of the 72 CSF samples (35%) was isolated. When comparing the cases group with control group, the OR of drinking home-made beverages was 4.1 (95% CI: 1.4-12.0), especially the beverages sacked by plastic bag: 3.3 (95% CI: 1.3-8.8). 6 out of 7 workers engaging in producing home-made beverages were detected to have carried Echovirus type 6 from their stool specimens. The Echovirus type 6 neutralization antibody positive rate in cases (73.5%) was significantly higher than that in 100 healthy people (46.0%) (X2 = 12. 526, P = 0.000).

CONCLUSIONThis episode of meningitis outbreak was caused by Echovirus type 6. The proportion of drinking home-made beverages, especially the beverages sacked by plastic bag in cases group was higher than in control group.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Echovirus 6, Human ; pathogenicity ; Echovirus Infections ; epidemiology ; etiology ; pathology ; virology ; Female ; Humans ; Male ; Meningitis ; epidemiology ; etiology ; pathology ; virology ; Prejudice

Echovirus 6 (ECV6) is the prevalent serotype detected in aseptic meningitis cases in Korea. To analyze the genetic variation of ECV6 isolates recently circulating in Korea, we determined the partial sequence of the VP1 capsid gene from 22 Korean ECV6 isolates and performed pairwise analysis against 42 reference strains from the GenBank database using MegAlign. The 22 Korean ECV6 isolates formed 3 distinct genetic clusters: Kor-lineage I, II, and III. The Korean ECV6 strains showed significant genetic diversity with 14.8~22.8% nucleotide divergence among the 3 different lineages. These ECV6 Kor-lineages were demonstrated to belong to different genetic clusters using VP1 sequence-based phylogenetic analysis, implying that the recently circulating Korean ECV6 strains have potential antigenic variation.
Antigenic Variation ; Capsid ; Databases, Nucleic Acid ; Echovirus 6, Human ; Enterovirus B, Human ; Genetic Variation ; Korea ; Meningitis, Aseptic

PURPOSE: There occurred an explosive outbreak of aseptic meningitis in Ulsan, in the year of 2002. In this study, we report clinical manifestations, laboratory data and causative viruses of the outbreak. METHODS: Were assessed 371 children with aseptic meningitis who were admitted to the Department of Pediatrics, Ulsan University Hospital in Ulsan between May and July, 2002. We reviewed the sex, age, clinical manifestations, peripheral blood examinations and CSF examinations of the patients. Virus isolation and serotype identification were performed by cell cultures of cerebrospinal fluid and stool. RESULTS: The ratio of males to females was 1.9:1. The majority of aseptic meningitis occurred in children between 1 and 5 years of age with a mean age of 6.8+/-5.1 years. The clinical manifestations was fever, headache, vomiting, and abdominal pain and 16.2% of the patients developed neck stiffness. The mean duration of fever was 3.2 days. The mean value of peripheral blood leukocytes was 10,355+/-4,800/mm3 and the mean value of CRP was 0.96+/-1.67 mg/dL. Initial CSF findings revealed leukocytes 212+/-385/mm3, protein 37.0+/-18.9 mg/dL and glucose 68.3+/-17.6 mg/dL. Viruses were isolated 67(18.1%) out of 371 patients and the serotypes were echovirus 6 in 10 cases, echovirus 9 in 15 cases, echovirus 11 in 4 cases, echovirus 13 in 18 cases, and unclassified enterovirus in 20 cases. CONCLUSION: Aseptic meningitis was prevalent in Ulsan, in the year of 2002. The clinical manifestations and the laboratory findings of the patients were compatible with those of previous reports. The causative viruses were echovirus 6, echovirus 9, echovirus 11, echovirus 13 and unclassified enterovirus.
Abdominal Pain ; Cell Culture Techniques ; Cerebrospinal Fluid ; Child ; Echovirus 6, Human ; Echovirus 9 ; Enterovirus ; Enterovirus B, Human ; Female ; Fever ; Glucose ; Headache ; Humans ; Leukocytes ; Male ; Meningitis, Aseptic* ; Neck ; Pediatrics ; Ulsan* ; Vomiting

PURPOSE: There was a outbreak of aseptic meningitis in Busan, 2002. We report the clinical features and causative viruses. METHODS: Two hundred seventy six children with aseptic meningitis who were admitted to the Department of Pediatrics, Maryknoll Hospital between January and December, 2002 were included. CSF, stool and throat swab viral cultures were done in 244 of these children. RESULTS: The male to female ratio was 1.4 : 1. Age of patients varied from five months to fourteen years old. Average age was 5.7+/-6.2 years old and mostly between one and four years.(41.3%) Monthly distribution revealed that the number of patients increased from May to July. The main symptoms were fever, headache and vomiting in this order. Respiratory tract infection symptom was associated from June to July and headache and nausea without fever were characteristically observed in children more than 10 years old from November to December. In peripheral blood examination, leukocytosis(WBC>10,000/mm3) showed in 34.8%, ESR was increased in 56.1%, and CRP was positive value in 61%. Therefore differential diagnosis was difficult through peripheral blood examination. CSF findings revealed mean leukocyte count 86.5+/-180.2/mm3, protein 41.7+/-32.9 mg/dL, glucose 56.4+/-9.9 mg/dL. Median hospitalized period was 4.7+/-7.2 days and compared with non-tapping group, hospitalized period was shorter and subsidance of symptoms was faster, therefore antibiotics injection period was shorter in the spinal tapping group. Virus was isolated in 31 cases of 244. The causative agents were echovirus 6, echovirus 9, echovirus 25, coxsakie virus B3, B4. CONCLUSION: There was an epidemics of aseptic meningitis in Busan, 2002; the causative agent was echovirus 6, 9, 25, coxsakie virus B3, B4.
Anti-Bacterial Agents ; Busan* ; Child ; Diagnosis, Differential ; Echovirus 6, Human ; Echovirus 9 ; Enterovirus B, Human ; Female ; Fever ; Glucose ; Headache ; Humans ; Leukocyte Count ; Male ; Meningitis, Aseptic* ; Nausea ; Pediatrics ; Pharynx ; Respiratory Tract Infections ; Spinal Puncture ; Vomiting

BACKGROUND: The determination of serotype of enteroviruses is useful for the discrimination between sporadic and epidemic infections. The conventional serotyping method is time-consuming and labor-intensive. Recently, molecular method was introduced for the serotyping of enteroviruses. The aim of this study was to establish a method to isolate and analyze enteroviruses from various specimens utilizing molecular biological techniques and to determine which strains were phylogenetically related to clinical samples. METHODS: Clinical samples in this study included 164 cerebrospinal fluid (CSF), 136 stool, 15 sera, 6 throat swab, 5 urine, and 4 sputa, which were obtained from hospitalized patients, primarily infants or children presenting symptoms of aseptic meningitis in 1998. RD cells were used for enterovirus isolation. RT-PCR was performed with RD cell lysate showing CPE. The primers 011 and 012 were used for the VP1 region, and the primers EN1 and EN2 for 5'-UTR. The nucleotide sequences of VP1 region were determined and analyzed with BLAST program. RESULTS: Among 333 samples, only 23 samples produced CPE: 17 samples at first and six samples at the second blind passage. Fifteen isolates were related to coxsackievirus B2 two to echovirus 4, three to echovirus 6, and three to echovirus 18. All 23 viral isolates displayed a nucleotide sequence identity of 80-95%, compared with the reference serotypes. However, the identity was increased up to 93-100% when the VP1 region was translated into amino acids CONCLUSIONS: Since CB2 type was 55% among enteroviral isolates, the CB2 was determined as the major causative serotype of enteroviral meningitis in 1998. CB2 type was emerged between June and July, EC4 and EC6 was limited to July, and EC18 was in August.
Amino Acids ; Base Sequence ; Cerebrospinal Fluid ; Child ; Discrimination (Psychology) ; Echovirus 6, Human ; Enterovirus B, Human ; Enterovirus* ; Humans ; Infant ; Meningitis* ; Meningitis, Aseptic ; Pharynx ; Serotyping*

BACKGROUND: The determination of serotype of enteroviruses is useful for the discrimination between sporadic and epidemic infections. The conventional serotyping method is time-consuming and labor-intensive. Recently, molecular method was introduced for the serotyping of enteroviruses. The aim of this study was to establish a method to isolate and analyze enteroviruses from various specimens utilizing molecular biological techniques and to determine which strains were phylogenetically related to clinical samples. METHODS: Clinical samples in this study included 164 cerebrospinal fluid (CSF), 136 stool, 15 sera, 6 throat swab, 5 urine, and 4 sputa, which were obtained from hospitalized patients, primarily infants or children presenting symptoms of aseptic meningitis in 1998. RD cells were used for enterovirus isolation. RT-PCR was performed with RD cell lysate showing CPE. The primers 011 and 012 were used for the VP1 region, and the primers EN1 and EN2 for 5'-UTR. The nucleotide sequences of VP1 region were determined and analyzed with BLAST program. RESULTS: Among 333 samples, only 23 samples produced CPE: 17 samples at first and six samples at the second blind passage. Fifteen isolates were related to coxsackievirus B2 two to echovirus 4, three to echovirus 6, and three to echovirus 18. All 23 viral isolates displayed a nucleotide sequence identity of 80-95%, compared with the reference serotypes. However, the identity was increased up to 93-100% when the VP1 region was translated into amino acids CONCLUSIONS: Since CB2 type was 55% among enteroviral isolates, the CB2 was determined as the major causative serotype of enteroviral meningitis in 1998. CB2 type was emerged between June and July, EC4 and EC6 was limited to July, and EC18 was in August.
Amino Acids ; Base Sequence ; Cerebrospinal Fluid ; Child ; Discrimination (Psychology) ; Echovirus 6, Human ; Enterovirus B, Human ; Enterovirus* ; Humans ; Infant ; Meningitis* ; Meningitis, Aseptic ; Pharynx ; Serotyping*