• Pediatric Infection and Vaccine
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• v.5 no.1
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• pp.104-114
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• 1998

Purpose : Enteroviruses are the most common cause of aseptic meningitis. The epidemics of aseptic meningitis in 1993 and 1996 were mostly caused by echovirus type 9. Identification of the causative virus of aseptic meningitis in epidemics, is very important not only for diagnosis but also for epidemiologic purpose. The purpose of this study was to identify the causative virus and investigate the relationship between aseptic meningitis, prevailed in Masan and surrounding areas in Kyoungsangnamdo in 1997, and its clinical manifestations. Methods : One hundred twenty eight cerebrospinal fluid(CSF) and 239 stool specimens were obtained from 239 patients(213 children and 26 adult patients) with aseptic meningitis were admitted to Masan Fatima Hospitals from March to October 1997. Viral isolation and serotype identification was performed by cell culture and immunofluorescent test. Enteroviruses not typed by immunofluorescent test was confirmed by reverse transcription-polymerase chain reaction(RT-PCR). Results : 1) The peak incidence was noted in June. 2) The age of 239 patients(pediatrics-213 cases, internal medicine-26 cases) that were diagnosed ranged from neonate to 35 years, the age of the patients of pediatrics ranged from neonate to 15years(mean 4.9 years), the age of the patients of internal medicine (above 16 years) ranged from 16 years to 35 years(mean 24.2 years). 3) Fifty-three(41.4%) of 128 CSF specimens were positive for enteroviruses, and 163(68.2%) of 239 stool specimens were positive for enteroviruses respectively. 4) Serotypes of 53 enteroviruses isolated from CSF were 16(30.2%) of echovirus type 30, 6(11.3%) of echovirus type 6, 1 of echovirus type 4, 4 of untyped echovirus, 1 of coxsackievirus type B5, and 24 isolates of untyped enteroviruses. Of 163 enterovirus isolated from stool were 72(44.2%) of echovirus type 30, 21(12.9%) of echovirus type 6, 1 of echovirus type 4, 17(10.4%) of undetermined subtyped echovirus, 1 of coxsackievirus type B5, 2 of A24, 3 of undetermined subtyped coxsackievirus type B, and 46 isolates of untyped enterovirus. Conclusion : There were epidemics of aseptic meningitis in the central areas of Kyoungsangnamdo from March to October 1997. The main causative organism was thought to be the echovirus type 30, and echovirus type 4, 6, coxsackievirus B5 and A24 were also thought to contribute to the epidemics.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.179-183
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• 2004

Enterovirus isolation was attempted from samples obtained from aseptic meningitis-suspected patients in hospitals in Busan during 2002. Enteroviruses were found in 83 of 703 cases. Echovirus serotypes 2, 3, 6, 7, 9, 13, 25, 29, 30 were isolated in 72 cases while coxsackievirus serotypes B3 and B4 were isolated in 11 cases. The occurrence was found to be distributed from April thru November with the highest rate during June and July. The strains of Vero and HEp-2 of echovirus and coxsackievirus, respectively, are highly infectious. Age distribution of patients, 61 patients in Enteroviruses and 11 patients in coxsackievirus, respectively, occurred in 0 to 10 year-olds. The sex distribution of the patients is as follows, 52 males (62.7％), 31 females (37.3％). Occurrence rate was found to be higher in male patients. Electron micrograph of echovirus and coxsackievirus show that they are small nonenevolped, isometric-shaped viruses. Isolated RNA from strains of echovirus and coxsackievirus showing cytopathic effects were used to undergo nested PCR which resulted in a 437 bp single band in all the strains.

• Korean Journal of Microbiology
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• v.39 no.4
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• pp.248-252
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• 2003

Enteroviruses isolation were attempted from samples obtained from aseptic meningitis-suspected patients in hospitals in Busan during 2000-2002. Enteroviruses were found in 2 of 292 cases in 2000, 4 of 371 cases in 2001, 83 of 703 cases in 2002. In 2000, the isolated viruses were found to be echovirus serotype 11 and coxsackievirus serotype B2. Coxsackievirus serotype B5 was isolated in 2001 and in 2002, echovirus serotypes 2, 3, 6, 7, 9, 13, 25, 30 were isolated in 70 cases while coxsackievirus serotypes B3 and B4 were isolated in 10 cases. Various specimens tended to emerge over the years. The occurrence in 2000 tended to be mostly focus during the cold months, December through January, while in 2001, it occurred in May. In 2002, occurrence was found to be distributed from April to November with the highest rate during June and July. The strains of Vero and HEp-2 of echovirus and coxsackievirus, respectively, are highly infectious. Electron micrograph of echovirus and coxsackievirus show that they are small nonenevolped, isometric-shaped viruses. Isolated RNA from strains of echovirus and coxsackievirus showing cytopathic effects were used to undergo nested PCR which resulted in a 436 bp single band in all the strains. The serotype was sent to the Department of Virology at the Korean National Institute of Health for identification.

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.289-293
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• 2001

Incidence of infectious viruses is ensuing throughout the world and threatening the health of children as well as adults. The outbreaks of viral diseases of alimentary tract in Pusan from 1998 to 2000 were detected. Viruses were isolated from stool specimens, cerebrospinal fluid and throat swabs from suspicious patients and confirmed by cell culture, latex agglutination test, indirect immunofluorescent test and electron microscopic observation. The average isolation rate was 12.5% from the suspected specimens. From this work, 2 cases of enteric adenoviruses, 23 cases of echovirus, 31 cases of coxsackivirus 36 cases of rotavirus, 45 cases of SRSV, and 7 cases of poliovirus were detected. The major serotypes of coxsackievirus were B2, B3, B4, B6 and echovirus of serotypes 6, 9, 11, 25, and 30 were examined. Two cases of enteric adenovirus type 41 were also confirmed. The incidence of SRSV was mostly concentrated between December through following March, April through October with echovirus and coxsackievirus, and January through April with rotavirus, respectively. Electron micrograph of negative-stained viruses showed typical appearance with 30-80 nm in diameter.

• Pediatric Infection and Vaccine
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• v.6 no.1
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• pp.123-130
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• 1999

Purpose : Meningitis is an inflammation of meninges by various kinds of organisms. Almost 85% of aseptic meningitis is caused by Enterovirus. This study was done to detect the causative virus of those with aseptic meningitis through sequencing the 5'-noncoding region to compare prototype and homology. Methods : RNA was extracted from Coxsackie Bl, Echovirus 3, 7, 9, 30. DNA was synthesized by RT-PCR and we compared homology with prototype from WHO by direct sequencing. Results : 1) PCR products from these viruses showed same bands of 155 bp and 440 bp on gel electrophoresis. 2) Coxsackievirus and Echovirus 11 prototype sequences were compared, which showed 12 bp changes with 92.1%. 3) Coxsackievirus B1 from a patient showed 94.1% homology when compared with prototype. 4) Echovirus 3 showed 92.8%, echovirus 7 92.8%, echovirus 30 82.9% homology. Conclusion : 5'-NCR of enterovirus has high homology which was good for use of diagnosis and more long sequencing requires for typing of viruses.

• Pediatric Infection and Vaccine
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• v.11 no.2
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• pp.176-182
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• 2004

Purpose : The etiologic agents of aseptic meningitis remain mostly unknown due to difficulty of viral culture and identification. There was an outbreak of aseptic meningitis in northern area of Seoul from June to August, 2002. We report the clinical features, laboratory data and causative viruses on 196 children with aseptic meningitis during this period. Methods : We retrospectively studied about clinical manifestations and laboratory findings 196 patients diagnosed as aseptic meningitis at Sanggye-Paik hospital. Virus isolation and serotype identification were performed by cell culture and reverse transcription polymerase chain reaction(RT-PCR) of the cerebrospinal fluid. Results : The male to female ratio was 1.39 : 1 and the mean age was 5.8+3 years. The clinical manifestations were fever, headache and vomiting. It occurred mostly in June, July and August. The numbers of peripheral blood leukocytes were $4,800{\sim}24,360/mm^3$. On cerebrospinal fluid examinations, leukocytes were in range of 10~2,000(mean 105)/$mm^3$, protein level in range of 15~171(mean 41.4) mg/dL and glucose level from 16~97(mean 57.9) mg/dL. Viral culture of cerebrospinal fluid showed 3 cases of Echovirus 9, 1 case of 25 and 30. In stool culture, 2 cases of Echovirus 6, 2 cases of Echovirus 13 and 1 case of Echovirus 30 were isolated. Conclusion : The etiologic viruses of the aseptic meningitis in northern area of Seoul in 2002 are presumed to be Echovirus 6, 9, 13, 25, 30.

• Pediatric Infection and Vaccine
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• v.20 no.2
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• pp.81-88
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• 2013

Purpose : This study was performed to investigate the epidemiology of enterovirus (EV) infections in children at a secondary hospital during recent 5 years. Methods : We collected the cerebrospinal fluid, stool and throat swab samples from the pediatric patients with suspected EV infections in KEPCO Medical Center, Seoul, Korea from July 2006 to September 2010. EV detection and genotype identification were performed by RT-PCR at Korea Centers for Disease Control and Prevention. Results : A total of 386 samples were collected from 277 patients during study period. Ninety-eight patients (35.4%) were diagnosed with EV infections. The RT-PCR positive rate was the highest in throat swab samples (48.3%). The median age of patient was 4.7 years (range, 0.1-12.5 years). Aseptic meningitis (50, 51.0%) was the most common clinical manifestation; herpangina (22, 22.4%) and hand-foot-mouth disease (18, 18.4%). One hundred EVs were isolated from 98 patients and 20 genotypes of EV were identified; Echovirus 30 (28 cases, 28%), Enterovirus 71 (12 cases, 12%), Echovirus 25 (10 cases, 10%), Echovirus 9 (9 cases, 9%) and Coxsackievirus A6 (8 cases, 8%). Aseptic meningitis caused by Echovirus 30 was the most common manifestation in 2008. There was no complicated case caused by Enterovirus 71. Conclusion : This study showed the epidemiology of confirmed EV infection in children from 2006 to 2010. There is a need for continuous surveillance of EV infections and its clinical manifestations.

• Journal of Environmental Science International
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• v.8 no.2
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• pp.165-169
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• 1999

The incidence of aseptic meningitis infection is ensuing and threatening the health of children. Enteroviruses are the major agents of aseptic meningitis and identification of virus has been a clue to diagnosis and epidemiology. The outbreak of aseptic meningitis occurred in Pusan, 1998. Patients were concentrated from April through November. Children were more susceptible than adults. Among 306 cases of specimens from stool, throat swab tested, only 7.2% were positive on virus isolation, 12 cases from stool and 10 from throat, respectively. All isolated 7 serotypes of viruses represented cytopathic effect on cultured cells. Three types of echovirus 6.25, 30 and coxsackievirus B2, B3, B4, B6 were identified by neutralizing antibody test. Isolated coxsackievirus and echovirus were observed by an electron microscope with negative staining.

• Clinical and Experimental Pediatrics
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• v.46 no.9
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• pp.858-864
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• 2003

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 $5.7{\pm}6.2years$ 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>$10,000/mm^3$) 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 $86.5{\pm}180.2/mm^3$, protein $41.7{\pm}32.9mg/dL$, glucose $56.4{\pm}9.9mg/dL$. Median hospitalized period was $4.7{\pm}7.2days$ 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.

• Pediatric Infection and Vaccine
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• v.5 no.1
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• pp.115-120
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• 1998

Purpose : In the summer 1997, there was an outbreak of aseptic meningitis in Pusan area. We report the clinical features and viral studies of aseptic meningitis in Pusan area in 1997. Methods : 265 children with aseptic meningitis who had been admitted to Department of Pediatrics, St. Benedict Hospital between April to October 1997 were included. Results : 1) Male-to-female ratio was 1.7:1. 2) Mean age was $6.2{\pm}3.29$ years. 3) It occured mostly April to October. 4) Clinical manifestations were fever 99.6%, vomiting 99.2%, headache 99.2%, rash 6.0%. 5) The duration of fever was $3.34{\pm}2.21$ days. 6) The duration of admission was $5.3{\pm}3.21$ days. 7) WBC count in peripheral blood were $11,200{\pm}4,163/mm^3$. 8) WBC count in CSF were $156.1{\pm}394.7/mm^3$. 9) Causative agents were coxsackievirus B5, echovius 6, 30, type nonspecific enterovirus. Conclusion : Aseptic meningitis in 1997 compared with that in 1996 had clinical feature of increase in age, decrease in duration of fever and incidende of rash. It occurred mostly April to October in 1997 and May to October in 1996. Causative agents were coxackievirus B5, echovirus 6, 30, type nonespecific enterovirus in 1997, and echovirus 9, coxsackievirus A24, type nonespecific enterovirus in 1996.