Carnobacterium Isolated from Caviar of Sturgeon (Acipenser ruthenus) Farmed in Korea

Sturgeon aqua-cultured in Korea is mainly Acipenser ruthenus and its culture began in the early 2000's. In this study, Carnobacterium sp. was isolated from unprocessed caviar of aqua-cultured Acipenser ruthenus. The 16s rRNA nucleotide sequence obtained from Carnobacterium sp. isolate (accession no. KM236206) was deposited with GenBank and homologous with Carnobacterium divergens DSM 20623 and NBRC15683 strain. In conclusion, this is first report of isolation of Carnobacterium sp. from caviar of Acipenser ruthenus aqua-cultured in Korea. In the future, it must be ascertained whether Carnobacterium sp. degenerate of caviar or cause diseases in sturgeon.

Antiviral Activity of Chrysin Derivatives against Coxsackievirus B3 in vitro and in vivo

Chrysin is a 5,7-dihydroxyflavone and was recently shown to potently inhibit enterovirus 71 (EV71) by suppressing viral 3C protease (3Cpro) activity. In the current study, we investigated whether chrysin also shows antiviral activity against coxsackievirus B3 (CVB3), which belongs to the same genus (Enterovirus) as EV71, and assessed its ability to prevent the resulting acute pancreatitis and myocarditis. We found that chrysin showed antiviral activity against CVB3 at 10 muM, but exhibited mild cellular cytotoxicity at 50 muM, prompting us to synthesize derivatives of chrysin to increase the antiviral activity and reduce its cytotoxicity. Among four 4-substituted benzyl derivatives derived from C(5) benzyl-protected derivatives 7, 9-11 had significant antiviral activity and showed the most potent activity against CVB3 with low cytotoxicity in Vero cells. Intraperitoneal injection of CVB3 in BALB/c mice with 1x106 TCID50 (50% tissue culture infective dose) of CVB3 induced acute pancreatitis with ablation of acinar cells and increased serum CXCL1 levels, whereas the daily administration of 9 for 5 days significantly alleviated the pancreatic inflammation and reduced the elevation in serum CXCL1 levels. Collectively, we assessed the anti-CVB3 activities of chrysin and its derivatives, and found that among 4-substituted benzyl derivatives, 9 exhibited the highest activity against CVB3 in vivo, and protected mice from CVB3-induced pancreatic damage, simultaneously lowering serum CXCL1 levels.

Genetic Characterization of Norovirus GII.3 Circulating in Korea

Noroviruses (NoV) are the major viral pathogen that causes epidemic acute gastroenteritis and outbreaks of food-borne illness. The major genotypes responsible for the epidemics of NoV are GII.4 and GII.3. However, most studies of NoV have been associated with GII.4 genotype and only few studies have been done with GII.3 genotype. Here, we selected 18 GII.3 strains, which recently circulated in Korea, and determined the partial sequences of the capsid gene. Phylogenetic analysis comparing these sequences with 29 reference strains from the GenBank database was performed using the MEGA program. All NoV GII.3 strains formed 2 distinct genetic lineages and variant groups. Lineage A showed 94.1~97.6%, 90.2~94.6% nucleotide identities from lineage B and variant group, respectively. Lineage B showed 90.2~94.6% nucleotide identities from variant group. These different genetic lineages based on the phylogenetic analysis of capsid sequences imply that the circulating Korean NoV GII.3 strains have potential antigenic variation.

Genetic Diversity of Echovirus 6 Strains Circulating in Korea

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.

Application of a Diagnostic Method Using Reverse Transcription-PCR ELISA for the Diagnosis of Enteroviral Infections / 대한진단검사의학회지

BACKGROUND: Enteroviruses are known as major pathogen for aseptic meningitis. Although rapid diagnosis for enteroviruses is very essential to exclude bacterial infections in patients with meningitis, classical diagnostic method based on virus isolation is not practicable for timely treatment of patients due to its laborious and time-consuming procedure. Recently molecular methodologies as alternatives are routinely used for rapid and sensitive diagnosis for enteroviruses infections. METHODS: Reverse transcription (RT)-PCR ELISA kit for targeting 5'non-coding region (NCR) with highly conserved genetic identity among all genotypes of enteroviruses was introduced in this investigation. RT-PCR ELISA was evaluated about sensitivity and specificity through virus isolation using clinical specimens from patients suspected of enteroviral infections and enteroviral isolates comparing with conventional RT-PCR identifying them. RESULTS: The detection limit of the RT-PCR ELISA was up to 10-100 folds higher than virus isolation using cell culture and conventional RT-PCR. On comparison between above two methods, the detection rate of RT-PCR ELISA for clinical specimens from patients with aseptic meningitis was 7% higher than that of conventional RT-PCR targeting 5'NCR (P=0.016). CONCLUSIONS: Our results suggest that RT-PCR ELISA developed in this study could be an alternative diagnostic method for the detection of enteroviral genome with high sensitivity and specificity.

Molecular identification and clinical features of enteroviral infection in children of central Korea: An overview of enteroviral epidemiology between spring 2005 and autumn 2006 / 소아과

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.

Molecular identification and clinical features of enteroviral infection in children of central Korea: An overview of enteroviral epidemiology between spring 2005 and autumn 2006 / 소아과

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.

Comparison of Nasopharyngeal Aspirates and Nasal Swabs for the Detection of Respiratory Viruses by Multiplex RT-PCR / 소아알레르기및호흡기학회지

PURPOSE: Although the nasopharyngeal aspirate (NPA) is more commonly used because of relatively higher accuracy, the nasal swab (NS) is a less painful and easier method than NPA. A few recent reports have shown that NS is more effective than NPA for the detection of respiratory virus using immunofluorescence or viral culture. The objective of the present study was to compare the results of NPA and NS sampling specimens in children for respiratory viruses detection using multiplex RT-PCR. METHODS: From December 2008 to June 2009 Paired NPA and NS specimens were collected from 250 children admitted with symptoms of acute respiratory infections at the Department of Pediatrics, Soonchunhyang University Cheonan Hospital, Cheonan, Korea. The sensitivity and agreement of virus detection between NPA and NS using multiplex RT-PCR were compared and analyzed. RESULTS: The median age of the subjects was 1.3 years (range, 20 days to 16.5 years), and 228 patients (91.2%) were under the age of 5 years. The agreement of virus detection between NPA and NS was excellent (Cohen's kappa >0.8) for parainfluenza virus type 3 or substantial (0.6 to 0.8) for rhinovirus A, RSV A and RSV B, moderate (0.4 to 0.6) for adenovirus and metapneumovirus and poor (<0.4) for influenza A. The overall sensitivity of detection of respiratory viruses was significantly higher in NPA (0.96) than in NS (0.59, P<0.05). CONCLUSION: We recommend NPA may be more accurate specimen for detection of respiratory viruses in hospitalized children. NS might be used in limited cases at a office setting or for larger epidemiological studies. However, results obtained from NS for influenza virus type A, metapneumovirus and adenovirus, should be interpreted carefully.

Comparison of Nasopharyngeal Aspirates and Nasal Swabs for the Detection of Respiratory Viruses by Multiplex RT-PCR / 소아알레르기및호흡기학회지

PURPOSE: Although the nasopharyngeal aspirate (NPA) is more commonly used because of relatively higher accuracy, the nasal swab (NS) is a less painful and easier method than NPA. A few recent reports have shown that NS is more effective than NPA for the detection of respiratory virus using immunofluorescence or viral culture. The objective of the present study was to compare the results of NPA and NS sampling specimens in children for respiratory viruses detection using multiplex RT-PCR. METHODS: From December 2008 to June 2009 Paired NPA and NS specimens were collected from 250 children admitted with symptoms of acute respiratory infections at the Department of Pediatrics, Soonchunhyang University Cheonan Hospital, Cheonan, Korea. The sensitivity and agreement of virus detection between NPA and NS using multiplex RT-PCR were compared and analyzed. RESULTS: The median age of the subjects was 1.3 years (range, 20 days to 16.5 years), and 228 patients (91.2%) were under the age of 5 years. The agreement of virus detection between NPA and NS was excellent (Cohen's kappa >0.8) for parainfluenza virus type 3 or substantial (0.6 to 0.8) for rhinovirus A, RSV A and RSV B, moderate (0.4 to 0.6) for adenovirus and metapneumovirus and poor (<0.4) for influenza A. The overall sensitivity of detection of respiratory viruses was significantly higher in NPA (0.96) than in NS (0.59, P<0.05). CONCLUSION: We recommend NPA may be more accurate specimen for detection of respiratory viruses in hospitalized children. NS might be used in limited cases at a office setting or for larger epidemiological studies. However, results obtained from NS for influenza virus type A, metapneumovirus and adenovirus, should be interpreted carefully.

Updates on Enterovirus Surveillance in Korea / 감염과화학요법

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.

Updates on Enterovirus Surveillance in Korea / 감염과화학요법

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.