• The Journal of Korean Society of Virology
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• v.27 no.2
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• pp.169-176
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• 1997

In this study, the feasibility of identification and genotypic differentiation of enteroviruses was investigated by using nested reverse transcription-polymerase chain reaction (nested RT-PCR), single-stranded conformation polymorphism (SSCP), and restriction fragment length polymorphism (RFLP) techniques. Two hundred seventy-four clinical samples were assayed by both nested RT-PCR and tube culture method using MRC-5 and MK cells; 58 (86.6%) out of 67 enterovirus culture-positive samples contained enteroviral RNA. In addition, 114 (55.1%) of 207 samples from patients with suspected enteroviral CNS disease with negative viral cultures were positive by the nested RT-PCR. The nested RT-PCR products were genotyped by the SSCP method and the results were compared with serotypes. We could differentiate 6 subtypes, 3 of which are similar to coxsackievirus B3, B5, echovirus 11, plus 3 other subtypes. RFLP cleaved with Sty I, Bgl I, and Xmn I yielded characteristic patterns for each laboratory strains. This study demonstrates the usefulness of the RT-PCR for the rapid diagnosis of enterovirus infection and the potentials of the SSCP method for differentiation of enterovirus strains.

• Pediatric Infection and Vaccine
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• v.3 no.2
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• pp.163-167
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• 1996

Aseptic meningitis, the most common infection of the central nervous system, is an acute illness mostly caused by enteroviruses. Cerebrospinal fluid(CSF) has been used for the detection of enteroviral RNA but the detection has been mostly performed in a single CSF specimen obtained during the illness. A major objective was to evaluate the relation of sampling time to the recovery of enteroviral RNA in CSF. Thirty seven CSF specimens were obtained from 24 patients between May and August 1993, when an outbreak of asceptic meningitis by echovirus type 9 occurred. Enteroviral RNA in CSF was detected by polymerase chain reaction(PCR). Data about onset of symptom development were obtained by review of medical records. Enteroviral RNA was detected by PCR in 29 of 37 CSF specimens. PCR yielded positive results in 4 of 5 CSF specimens obtained on day 1 to 3, 10 of 11 on day 4 to 6, 8 of 10 on day 7 to 9, 6 of 8 on day 10 to 12, 1 of 3 on day 13 to 15 postonset. Of 11 patients from each of whom more than one CSF were obtained on different day postonset, PCR yielded positive resutls in 2 of 3 cases in whom enteroviral RNA detection was negative in the first CSF. These results indicate that two or more CSF specimens obtained within 12 days postonset are required for improving the accuracy of the diagnosis of enteroviral meningitis.

• Proceedings of the Microbiological Society of Korea Conference
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• 2000.05a
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• pp.94-101
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• 2000

We investigated the viral contamination of water environment including tap water in Korea. River water used for source water was analyzed about monthly between 1997 and 1999 over a period 26 months. A total of 22 tap water samples were collected in 10 sites in 2 urban areas between 1997 and 1998 over a 11 months. All samples were examined for infectious enteroviruses and adenoviruses by a cell culture technique followed by PCR amplification. To identify the recovered viruses from tap water, sequence analysis of PCR products was performed. Infectious viral particles were detected in river water all year round, ranging from 0.93 to 17.3 Most Probable Number of Infectious Unit (MPNIU) /100L. Tap water samples also contained infectious viral particles. The frequency of enteroviruses and adenoviruses in tap water were $50.0\%$ (11/22) and $36.7\%$ (8/22), respectively. Both enteroviruses and adenoviruses were detected in five tap water samples $(22.7\%)$. The level of viral contamination in tap water was quite high, ranging from 0.2 to 2.9 MPNIU/100L, far above the recommended virus level in drinking water set by the U.S. EPA. Poliovirus type 1 derived from vaccine was frequently detected and the remainder comprised coxsackievirus B type or echovirus type 6, which were causative agents of aseptic meningitis in Korea in 1997 and 1998, respectively. Several types of adenovirus were detected in tap water samples and some water samples were found to contain adenoviruses which were closely related to enteric adenovirus type 40 and 41. This stusy shows that surface water and tap water in Korea may be exposed to the risk of viral contamination, especially from recently recognized viruses and this constitutes a potential public health hazard.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.1055-1064
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• 2009

Enteroviruses were identified and characterized from patients with aseptic meningitis and other enterovirus-related diseases in Chungnam, Korea from 2005 to 2006. Enteroviruses were isolated from 79 of 519 cases (15.2%) in 2005, and 37 of 386 cases (9.6%) in 2006. Based on partial VP1 sequencing, a total of 116 enterovirus isolates were resolved into 13 types. Prevalent among the Chungnam isolates were echovirus 18 and coxsackievirus B5 in 2005, and echoviruses 5 and 25 in 2006. This is the first time echoviruses 5 and 18 have been identified in Korea since enterovirus surveillance began there in 1993. The temporal distribution of enterovirus epidemics in Chungnam showed a remarkable seasonal pattern, with cases occurring during most of the three months of the summer from June to August. The highest rate of enterovirus-positive cases occurred in patients less than 1 year of age. The ratio of male to female enterovirus-positive patients was approximately 1.8:1. Comparison of the VP1 amino acid sequences of the 15 coxsackievirus B5 isolates with reference strains revealed that all Chungnam isolates are substituted at positions 23 (V231), 19 (S19G), 75 (Y75F), and 95 (N95S). Upon comparing the nine ECV5 isolates with foreign strains, it was found that only the Chungnam isolates, with the exception of Kor06-ECV5-239cn, have P at position 153 and F at position 146. The three ECV9 isolates from 2006 show alterations at amino acids 36, 148, and 154 outside of the BC-loop and at position 84 in the BC-loop, whereas the seven isolates from 2005 and the other ECV9 strains in the database only show the alteration at position 84 (D, I, N, S). The five ECV25 isolates have an S residue at position 134, whereas most of the foreign strains have an N residue.

• Pediatric Infection and Vaccine
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• v.6 no.2
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• pp.210-218
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• 1999

Purpose : As several epidemics of aseptic meningitis had been reported in Korea since 1990, the need of epidemiologic study about enteroviral infections was recognized. We studied epidemiologic and clinical features of enteroviral infections in children. Methods : We analyzed the seasonal occurrence of enteroviral infections and age, sex and clinical diagnosis of the children admitted to five branch hospitals of Hallym University and diagnosed to have enteroviral infections by culture from January 1996 to December 1998. Results : Enterovirus was isolated in 126 out of 245(51.4%) stools, 15 out of 89(16.8%) nasopharyngeal aspirates and 195 out of 1,835(10.6%) cerebrospinal fluids. There were 273 echoviruses(Echo)(197 Echo 30, 46 Echo 9, 17 Echo 6, 13 unclassified), 24 coxsackieviruses B(CB)(11 CB 2, 2 CB 5, 11 unclassified), 7 coxsackieviruses A 24 and 32 unclassified enteroviruses. Yearly number of isolates was 65(46 Echo 9) in 1996, 239(197 Echo 30) in 1997 and 32(15 CB: 9 CB 2) in 1998. Epidemics of enteroviruses occurred mainly during the months May to October. The mean age of the infected children was $62.1{\pm}38.0$ months and 74.6% of the patients were younger than 7 years of age. There were 203 boys and 100 girls and 79.3% of the patients were diagnosed as aseptic meningitis. Conclusions : We confirmed that enteroviral infections occurred in epidemics from late spring to fall during the study period. Enteroviral infections occurred predominantly in boys and the preschool children. Aseptic meningitis was the main illness in the hospitalized children.

• Pediatric Infection and Vaccine
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• v.14 no.2
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• pp.179-187
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• 2007

Purpose : Acute viral respiratory tract infection is the most common illness among children. Involved organisms are respiratory syncytial virus (RSV), influenza virus, adenovirus, parainfluenza virus, etc. The objective of the present study is to determine epidemiology of each viral infection in Busan, South Korea between January 2004 and December 2006. Methods : We retrospectively analyzed the results of clinical samples of throat and nasal swab collected from Busan city, South Korea between January 2004 and December 2006. We collected these samples from the children with acute respiratory illness. Viruses were detected by virus culture Results : The identified pathogens were influenza A in 131 cases (46.6 percent), RSV in 40 cases (14.2 percent), influenza B in 37 cases (13.2 percent), adenovirus in 23 cases (8.2 percent), enterovirus in 37 cases (13.2%) [coxsackie virus in 20 cases (7.1 percent), atypical enterovirus in 9 cases (3.2 percent), poliovirus in 4 cases (1.4 percent), echovirus in 4 cases (1.4 percent)], rhinovirus in 9 cases (3.2 percent), parainfluenza virus in 2 cases (0.7 percent), coronavirus and bocavirus in each 1 case (0.4 percent). Influenza A were detected every year, mostly in December through April and RSV in October through February. Adenovirus were detected through out the year. Bocavirus occurred in December, 2006. Conclusion : We analyzed epidemiologic characteristics of viruses to cause the respiratory disease that prevailed at Busan area for recent three years, and from now on, a further continuous study will be necessary.