The environmental water samples assayed by total culturable virus assay (TCVA) were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and integrated cell culture-PCR (ICC-PCR) method for enteroviruses and reovirus since they are the usually detected virus groups by culture assays. The detection sensitivities of the TCVA, RT-PCR, and ICC-PCR were compared and the overall reliability of the detection was analyzed to confirm environmental samples for enteric viruses. A total of eight samples from different areas was analyzed by performing TCVA, RT-PCR, and ICC-PCR. Virus concentrations in surface water samples ranged from 1.03 to 47.3 most probable numbers of infectious units (MPN) per 100 liters. When primers specific for both enteroviruses and reoviruses were used in both RT-PCR and ICC-PCR, all the samples (100%) were positive for the viruses. Reoviruses were the most frequently detected ones among the samples. According to the sequence results of enteroviruses, five of the samples were contaminated by coxsackievirus type B3, and the rest by coxsackievirus type B6, echovirus type 30, or vaccine strain poliovirus type 3. It was observed that both enteroviruses and reoviruses were detected concurrently in all CPE-positive environmental water samples.
Enterovirus B, Human ; Enterovirus* ; Poliovirus ; Water*

The environmental water samples assayed by total culturable virus assay (TCVA) were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and integrated cell culture-PCR (ICC-PCR) method for enteroviruses and reovirus since they are the usually detected virus groups by culture assays. The detection sensitivities of the TCVA, RT-PCR, and ICC-PCR were compared and the overall reliability of the detection was analyzed to confirm environmental samples for enteric viruses. A total of eight samples from different areas was analyzed by performing TCVA, RT-PCR, and ICC-PCR. Virus concentrations in surface water samples ranged from 1.03 to 47.3 most probable numbers of infectious units (MPN) per 100 liters. When primers specific for both enteroviruses and reoviruses were used in both RT-PCR and ICC-PCR, all the samples (100%) were positive for the viruses. Reoviruses were the most frequently detected ones among the samples. According to the sequence results of enteroviruses, five of the samples were contaminated by coxsackievirus type B3, and the rest by coxsackievirus type B6, echovirus type 30, or vaccine strain poliovirus type 3. It was observed that both enteroviruses and reoviruses were detected concurrently in all CPE-positive environmental water samples.
Enterovirus B, Human ; Enterovirus* ; Poliovirus ; Water*

PURPOSE: Enteroviruses are the most common cause of aseptic meningitis in patients of all ages. A definite diagnosis of enteroviral meningitis can be established by detection of virus directly in CSF specimens. But this is time-consuming and lacks sensitivity, so polymerase chain reaction(PCR) detecting of viral RNA in patient specimens such as CSF, stool has been demonstrated. But little is known about the influence of sampling time on the results of CSF PCR and stool PCR. We investigated diagnostic utility of PCR of CSF and stool according to sampling time after the onset of symptoms. METHODS: PCR results were analyzed according to sampling time for 42 patients diagnosed aseptic meningits in our hospital from 11(th) January to 30(th) August, 2005. RESULTS: The diagnostic yield of the test was higher of CSF specimens obtained < or = 2 days after clinical onset(positive PCR results 9/18, 50 percent), compared with CSF collected >2 days after onset(positive PCR results 1/24, 4.2 percent)(P=0.001). Instead, positive PCR results of fecal specimens maintained highly(average 90.5 percent), 10 cases had also positive PCR results even 5-6 days after onset. 10 cases of CSF specimens had positive enterovirus PCR results containing coxsackievirus B5 (n=6), coxsackievirus B3(n=3). 38 cases of stool specimens had positive enterovirus PCR results containing echovirus 18(n=7), echovirus 9(n=3), coxsackievirus B5(n=8), coxsackievirus B3(n=3). 6 cases(coxackie B5) had positive CSF PCR and stool PCR, both. CONCLUSION: Stool PCR was clinically sensitive for detecting enterovirus during enteroviral meningits and could give a presumptive diagnosis throughout the disease course. A definite diagnosis was obtained by CSF PCR, but its utility was clearly lower for samples obtained >2 days after clinical onset. Therefore, it is recommended that, in addition to performance of CSF PCR, fecal samples obtained from patients with suspected enteroviral meningitis should be tested by PCR, especially when the duration of symptoms is >2 days.
Cerebrospinal Fluid ; Diagnosis ; Enterovirus ; Enterovirus B, Human ; Humans ; Meningitis ; Meningitis, Aseptic* ; Polymerase Chain Reaction ; RNA, Viral

Echovirus 30 is one of the distinct serotypes of enteroviruses and commonly isolated agent causing sporadic to large outbreaks with aseptic meningitis in many regions over the world. Recently, an outbreak of echovirus 30 associated with aseptic meningitis occurred in Korea in 2008. In order to analyze echovirus 30 in Korea, the virus was isolated from cerebrospinal fluid samples of a male patient with aseptic meningitis and its genome sequence was determined. The sequence of Korean echovirus 30 isolate was compared with those of reference strains (Bastianni, FDJS03-84, zhejiang-17-03, 14916net87). At the nucleotide level, the P1 region (84.8~89.0%) had the highest identity value; at the amino acid level, the P3 region (97.0~98.5%) showed the highest value. When the cleavage sites were compared, most sites were identical except those between VP1 and 2A; the Bastianni stain had TT/GA, whereas the other four strains contained NT/GA. The China strains (FDJS03-84 and zhejiang-17-03) were grouped together and the other strains were distinct from each branch in the phylogenetic tree based on the complete genome sequences.
China ; Disease Outbreaks ; Enterovirus ; Enterovirus B, Human ; Genome ; Humans ; Korea ; Male ; Meningitis, Aseptic ; Sequence Analysis ; Viruses

PURPOSE: Enteroviruses (EVs) are commonly known to cause infection, especially in infants and children. This report presents an overview of enterovirus epidemiology in central Korea. METHODS: From the spring of 2005 to the autumn of 2006, we collected the cerebrospinal fluid (CSF) and stool samples from the pediatric patients with a febrile illness or suspected meningitis who were admitted to hospitals in central Korea. In order to test for EVs, cell lines were derived from pretreated susceptible specimen, and the cytopathic effects were observed. Seminested real time-polymerase chain reaction (RT-PCR) and direct sequencing were performed for genotypic and phylogenetic analyses. RESULTS: Of the 305 patients examined, 51 (16.7%) tested positive for EV. Of these 51 patients, 44 showed the following serotypes: Echovirus (ECV) 18 (18 cases, 35.2%), Coxsackievirus B (CVB) 5 (13 cases, 25.4%), ECV25 (5 cases, 9.8%), ECV9 (4 cases, 7.8%), ECV5 (3 cases, 5.8%), and EV74 (1 case, 1.9%). In 2005, between June and August, ECV18 and CVB5 were mostly responsible for the enteroviral infections among the patients in central Korea. In 2006, between July and August, ECV25 was mostly the cause of enteroviral infection. Conclusions: There is a need for continuous surveillance of enteroviral infection and its clinical manifestations, particularly for EV74, which was first identified in Korea.
Cell Line ; Child ; Enterovirus ; Enterovirus B, Human ; Humans ; Infant ; Korea ; Meningitis

PURPOSE: Enteroviruses (EVs) are commonly known to cause infection, especially in infants and children. This report presents an overview of enterovirus epidemiology in central Korea. METHODS: From the spring of 2005 to the autumn of 2006, we collected the cerebrospinal fluid (CSF) and stool samples from the pediatric patients with a febrile illness or suspected meningitis who were admitted to hospitals in central Korea. In order to test for EVs, cell lines were derived from pretreated susceptible specimen, and the cytopathic effects were observed. Seminested real time-polymerase chain reaction (RT-PCR) and direct sequencing were performed for genotypic and phylogenetic analyses. RESULTS: Of the 305 patients examined, 51 (16.7%) tested positive for EV. Of these 51 patients, 44 showed the following serotypes: Echovirus (ECV) 18 (18 cases, 35.2%), Coxsackievirus B (CVB) 5 (13 cases, 25.4%), ECV25 (5 cases, 9.8%), ECV9 (4 cases, 7.8%), ECV5 (3 cases, 5.8%), and EV74 (1 case, 1.9%). In 2005, between June and August, ECV18 and CVB5 were mostly responsible for the enteroviral infections among the patients in central Korea. In 2006, between July and August, ECV25 was mostly the cause of enteroviral infection. Conclusions: There is a need for continuous surveillance of enteroviral infection and its clinical manifestations, particularly for EV74, which was first identified in Korea.
Cell Line ; Child ; Enterovirus ; Enterovirus B, Human ; Humans ; Infant ; Korea ; Meningitis

PURPOSE: Enterovirus infection in children can manifest various disease and enterovirus have many serotypes. This study was aimed to investigate neurologic manifestations according to serotypes of enterovirus in pediatric inpatients in Incheon. METHODS: We collected the stool samples from the admitted pediatric patients in Inha University Hospital from January 2015 to September 2016. Enterovirus detection and serotypes identification were performed by real-time reverse transcriptase polymerase chain reaction (RT-PCR) and semi-nested RT-PCR. RESULTS: A total of 527 samples were collected during study period and 170 patients (32.2%) were diagnosed with enterovirus infections. Genetic sequences of enteroviruses were identified: echovirus 18 (50, 40.5%), enterovirus 71 (12, 9.6%), coxakievirus A10 (10, 8.0%), echovirus 6 (7, 5.6%). Virus in patient with meningitis were identified: echovirus 18 (15, 75%), coxakievirus B5 (2, 10%), enterovirus 71 (2, 10%), and echovirus 6 (1, 5%). Neurologic manifestations of echovirus 18 are headache (15, 30%), vomiting (17, 34%), meningeal irritation sign (10, 20.0%). And enterovirus 71 have headache (3, 25%), vomiting (3, 25%), meningeal irritation sign (2, 16.0%), seizure (1, 8.3%), neurologic sequelae (1, 8.3%). Echovirus 18 and neurologic manifestation have a statistically significant correlation with other serotypes (r=0.701, P < 0.01) CONCLUSION: Echovirus 18 infection was more prominent in neurological symptoms than in other serotypes. The major serotype of meningitis was echovirus 18 but there was no reported neurologic sequelae. Enterovirus infection has different neurological symptoms, depending on the serotypes.
Child ; Echovirus 6, Human ; Enterovirus B, Human ; Enterovirus Infections ; Enterovirus* ; Headache ; Humans ; Incheon* ; Inpatients* ; Meningitis ; Neurologic Manifestations* ; Reverse Transcriptase Polymerase Chain Reaction ; Seizures ; Serogroup* ; Vomiting

PURPOSE: We identified the causative viruses from patients with aseptic meningitis, acute hemorrhagic conjunctivitis and other enterovirus-related diseases to understand the epidemiological patterns and prevailing strains of enterovirus infections each year. MATERIALS AND METHODS: During 1999-2003, we examined 3,260 specimens from 2,939 patients with aseptic meningitis or other clinical manifestations for the presence of enteroviruses by using both cell culture/ neutralisation test and reverse transcription-polymerse chain reaction-sequencing. To investigate the etiological agents which caused an epidemic of acute haemorrhagic conjunctivitis, conjunctival swab samples from acute haemorrhagic conjunctivitis patients showing cytopathic effects in HEp2 cells were tested by enteroviral specific PCR. RESULTS: We identified 603 isolates of enteroviruses (20.5%) among 2,939 cases and 22 serotypes of human enteroviruses were isolated during this 5 year period. Echovirus 13 and coxsackievirus A24 in 2002 and coxsackievirus A9 in 2003 were the first enterovirus to be indentified in Korea since we began the enterovirus surveillance in 1993. While an epidemic of echovirus 13 infection in Korea began in Gwangju and Jeolla province in 2002 and spread to Seoul, Gyunggi, Busan, Ulsan and other regions, echovirus 6 isolates in 2002 were mainly detected in Busan specimens and some Gwangju samples. From the nucleotide sequencing of enteroviral PCR products of conjunctival swab specimens, we found 85% nucleotide homology to coxsackievirus A24 (D90457). CONCLUSIONS: We isolated 603 enteroviral isolates among 2939 cases during 1999-2003. Echovirus 13 and coxsackievirus A24 were the first enterovirus to be identified in Korea and caused nationwide epidemics in 2002.
Busan ; Conjunctivitis ; Conjunctivitis, Acute Hemorrhagic ; Echovirus 6, Human ; Enterovirus B, Human ; Enterovirus Infections ; Enterovirus* ; Gwangju ; Humans ; Korea* ; Meningitis, Aseptic ; Polymerase Chain Reaction ; Seoul ; Ulsan

PURPOSE: We identified the causative viruses from patients with aseptic meningitis, acute hemorrhagic conjunctivitis and other enterovirus-related diseases to understand the epidemiological patterns and prevailing strains of enterovirus infections each year. MATERIALS AND METHODS: During 1999-2003, we examined 3,260 specimens from 2,939 patients with aseptic meningitis or other clinical manifestations for the presence of enteroviruses by using both cell culture/ neutralisation test and reverse transcription-polymerse chain reaction-sequencing. To investigate the etiological agents which caused an epidemic of acute haemorrhagic conjunctivitis, conjunctival swab samples from acute haemorrhagic conjunctivitis patients showing cytopathic effects in HEp2 cells were tested by enteroviral specific PCR. RESULTS: We identified 603 isolates of enteroviruses (20.5%) among 2,939 cases and 22 serotypes of human enteroviruses were isolated during this 5 year period. Echovirus 13 and coxsackievirus A24 in 2002 and coxsackievirus A9 in 2003 were the first enterovirus to be indentified in Korea since we began the enterovirus surveillance in 1993. While an epidemic of echovirus 13 infection in Korea began in Gwangju and Jeolla province in 2002 and spread to Seoul, Gyunggi, Busan, Ulsan and other regions, echovirus 6 isolates in 2002 were mainly detected in Busan specimens and some Gwangju samples. From the nucleotide sequencing of enteroviral PCR products of conjunctival swab specimens, we found 85% nucleotide homology to coxsackievirus A24 (D90457). CONCLUSIONS: We isolated 603 enteroviral isolates among 2939 cases during 1999-2003. Echovirus 13 and coxsackievirus A24 were the first enterovirus to be identified in Korea and caused nationwide epidemics in 2002.
Busan ; Conjunctivitis ; Conjunctivitis, Acute Hemorrhagic ; Echovirus 6, Human ; Enterovirus B, Human ; Enterovirus Infections ; Enterovirus* ; Gwangju ; Humans ; Korea* ; Meningitis, Aseptic ; Polymerase Chain Reaction ; Seoul ; Ulsan

Objective: To develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses (HEVs) from clinical samples and to contribute to etiological surveillance of HEV-related diseases. Methods: A panel of RT-nPCR assays, consisting of published combined primer pairs for VP1 genes of HEV A-C and in-house designed primers for HEV-D, was established in this study. The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID Results: The sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID Conclusion This RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A-D, providing rapid, sensitive, and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.
Capsid Proteins/genetics* ; Enterovirus A, Human/genetics* ; Enterovirus B, Human/genetics* ; Enterovirus C, Human/genetics* ; Enterovirus D, Human/genetics* ; Humans ; Molecular Epidemiology/methods* ; Molecular Typing/methods* ; Reverse Transcriptase Polymerase Chain Reaction/methods*