Study on the complete sequence of CA24 variant isolated during the acute hemorrhagic conjunctivitis outbreaks in Zhejiang province during 2002 to 2010 / 中华流行病学杂志

Objective To analyze the genetic characteristics of the complete sequence of coxsackievirus A24 variant(CA24v) isolated from acute hemorrhagic conjunctivitis (AHC) outbreaks in Zhejiang province during 2002 to 2010.Methods Complete sequences of CA24v epidemic strains isolated in different years were amplified under the RT-PCR assay,while the sequences of whole genome,VP1,and 3C region of Zhejiang strains were compared with epidemic strains isolated in other areas of China and abroad.Results The whole genome of Zhejiang CA24v strains isolated in 2002 and 2010 was 7456-7458 bp in length,encoding a polyglutamine protein which containing 2214 amino acid residues.There was a insertion with T on site 97 and 119 within 5' non-coding region between epidemic strain Zhejiang/08/10 and strains isolated in 2002.The rates of amino acid homology among Zhejiang/08/10 and other strains isolated since 2002 were between 94.7％ and 100.0％.Compared with the representative strains circulated within the recent 60 years,the largest average amino acid variations had been occurred on region 2A and 3A,with the ratios as 8.4％ and 7.3％ respectively.The smallest variation happened in region 3D,with the ratio only as 1.9％.The rates of stable amino acid variation on the whole genome between strains isolated since 1987 and 2002 were 38 and 20.P-distance within groups appeared that region 3C was more stable than VP1 of strains isolated in 2002-2010,and the 3D of early strain Jamaica/10628/87 might have had a nature of recombination but not observed on those epidemic strains in recent years.Conclusion Within the evolution of CA24v strains,the time course was more significant than the geographical differences.There had been sporadic epidemics of AHC caused by CA24v in Zhejiang province since 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.

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.

Detection and Phylogenetic Analysis of Coxsackievirus A24 Variant Causing Nation-wide Epidemic of Acute Hemorrhagic Conjunctivitis in Korea, 2002 / 감염과화학요법

BACKGROUND: Nation-wide outbreak of acute hemorrhagic conjunctivitis occurred in the summer, 2002 in South Korea. We identified the causative agent of this outbreak through virus culture and molecular biological techniques. METHODS: Polymerase chain reaction (PCR) was carried out with direct conjunctival swab samples and cell culture supernatants. Conjunctival swab was done at a community based-eye clinic in Seoul, September 2002. Initial screening for adenovirus and enterovirus was performed. Nested PCR for adenovirus was done with adenovirus common primers using direct swab sample, and reverse transcription PCR (RT-PCR) for enterovirus was done with enterovirus common primers. RT-PCR with primer 188/222 for VP1 region of enterovirus was done, if initial screening test was positive. PCR product was sequenced, and homology searching, compared to prototype strains, was done for serotyping. Protease 3C region of coxsackievirus A24v was amplified and sequenced with primer D1/U2. The sequence of this region was compared to those of viral isolates, which had been obtained from several Asian outbreaks since 1970. RESULTS: Conjunctival swabs were performed in 88 patients. Thirty nine (44%) samples out of the 88 were culture positive on HeLa or MRC-5 cells. Nine (100%) out of 9 culture supernatants, randomly selected from 39 culture positve samples, were positive for coxsackievirus A24v-specific RT-PCR. Phylogenetic analysis showed that sequences from 14 culture positive supernatants, randomly selected from 39 culture positive samples, clustered into a time-related, but distinct lineage, with Asian strains. CONCLUSIONS: We identified the causative agent of the epidemic hemorrhagic conjunctivits in year 2002 as coxsackievirus A24v.

Detection and Phylogenetic Analysis of Coxsackievirus A24 Variant Causing Nation-wide Epidemic of Acute Hemorrhagic Conjunctivitis in Korea, 2002 / 감염과화학요법

BACKGROUND: Nation-wide outbreak of acute hemorrhagic conjunctivitis occurred in the summer, 2002 in South Korea. We identified the causative agent of this outbreak through virus culture and molecular biological techniques. METHODS: Polymerase chain reaction (PCR) was carried out with direct conjunctival swab samples and cell culture supernatants. Conjunctival swab was done at a community based-eye clinic in Seoul, September 2002. Initial screening for adenovirus and enterovirus was performed. Nested PCR for adenovirus was done with adenovirus common primers using direct swab sample, and reverse transcription PCR (RT-PCR) for enterovirus was done with enterovirus common primers. RT-PCR with primer 188/222 for VP1 region of enterovirus was done, if initial screening test was positive. PCR product was sequenced, and homology searching, compared to prototype strains, was done for serotyping. Protease 3C region of coxsackievirus A24v was amplified and sequenced with primer D1/U2. The sequence of this region was compared to those of viral isolates, which had been obtained from several Asian outbreaks since 1970. RESULTS: Conjunctival swabs were performed in 88 patients. Thirty nine (44%) samples out of the 88 were culture positive on HeLa or MRC-5 cells. Nine (100%) out of 9 culture supernatants, randomly selected from 39 culture positve samples, were positive for coxsackievirus A24v-specific RT-PCR. Phylogenetic analysis showed that sequences from 14 culture positive supernatants, randomly selected from 39 culture positive samples, clustered into a time-related, but distinct lineage, with Asian strains. CONCLUSIONS: We identified the causative agent of the epidemic hemorrhagic conjunctivits in year 2002 as coxsackievirus A24v.