Coxsackievirus A16 belongs to the family Picornaviridae, and is a major agent of hand-foot-and-mouth disease that infects mostly children, and to date no vaccines or antiviral therapies are available. 2A protease of enterovirus is a nonstructural protein and possesses both self-cleavage activity and the ability to cleave the eukaryotic translation initiation factor 4G. Here we present the crystal structure of coxsackievirus A16 2A protease, which interestingly forms hexamers in crystal as well as in solution. This structure shows an open conformation, with its active site accessible, ready for substrate binding and cleavage activity. In conjunction with a previously reported "closed" state structure of human rhinovirus 2, we were able to develop a detailed hypothesis for the conformational conversion triggered by two "switcher" residues Glu88 and Tyr89 located within the bll2-cII loop. Substrate recognition assays revealed that amino acid residues P1', P2 and P4 are essential for substrate specificity, which was verified by our substrate binding model. In addition, we compared the in vitro cleavage efficiency of 2A proteases from coxsackievirus A16 and enterovirus 71 upon the same substrates by fluorescence resonance energy transfer (FRET), and observed higher protease activity of enterovirus 71 compared to that of coxsackievirus A16. In conclusion, our study shows an open conformation of coxsackievirus A16 2A protease and the underlying mechanisms for conformational conversion and substrate specificity. These new insights should facilitate the future rational design of efficient 2A protease inhibitors.
Coxsackievirus Infections ; virology ; Crystallography, X-Ray ; Cysteine Endopeptidases ; chemistry ; genetics ; Fluorescence Resonance Energy Transfer ; Hand, Foot and Mouth Disease ; enzymology ; pathology ; virology ; Humans ; Picornaviridae ; chemistry ; enzymology ; genetics ; Protein Conformation ; Structure-Activity Relationship ; Substrate Specificity ; Viral Proteins ; chemistry ; genetics

This article describes the nomenclature, history and genetic evolution of duck hepatitis A virus, and updates the epidemiology, clinical symptom and surveillances of duck virus hepatitis A. It also summarizes the present status and progress of duck virus hepatitis A and illustrated the necessity and urgency of its research, which provides rationale for the control of duck hepatitis A virus disease in China.
Animals ; Ducks ; virology ; Hepatitis Virus, Duck ; classification ; genetics ; isolation & purification ; Hepatitis, Viral, Animal ; virology ; Picornaviridae Infections ; veterinary ; virology

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

Both sides of the picornavirus genome have 5'-untranslated region (5'UTR) and 3'- untranslated region (3'UTR). This study demontrated that both the 5'-and 3'-UTR can form complex structures, such as stem-loop, clover and pseudoknot structure, These structures play an important role in the regulaton of the replication and translation of the viruses. This article reviewed the progress of research on the structure and function of picornavirus' 3'-UTR over recent years.
3' Untranslated Regions ; Animals ; Humans ; Nucleic Acid Conformation ; Picornaviridae ; chemistry ; genetics ; metabolism ; Picornaviridae Infections ; virology ; RNA, Viral ; chemistry ; genetics ; metabolism

The rapid mutation and widely spread of duck hepatitis A virus (DHAV) lead to the vast economic loss of the duck industry. To prepare and evaluate bivalent inactivated vaccine laboratory products of DHAV, 6 strains were screened from 201 DHAV-1 strains and 38 DHAV-3 strains by using serotype epidemiological analysis in most of the duck factory. Vaccine candidate strains were selected by ELD50 and LD50 tests in the 6 strains. Continuously passaged, the 5th passaged duck embryos bodies grinding fluid was selected as vaccine virus seeds. The virus seeds were treated with formaldehyde and water in oil in water (W/O/W) emulsions, making into three batches of two bivalent inactivated vaccine laboratory products. The safety test, antibody neutralization test, challenged protection and cross immune protection experiment suggested that the vaccines possessed good safety, and neutralizing antibodies were detected at 7th day and the challenged protection rate reached 90% to 100% at the 14th and 21st day. Moreover, immune duration of ducklings lasted more than five weeks. However, cross-immunity protection experiments with DHAV-SH and DHAV-FS only had 20%-30%. The two bivalent inactivated vaccine laboratory products of duck viral hepatitis were effective and reliable, providing a new method as well as a new product for DHAV prevention and control.
Animals ; Antibodies, Neutralizing ; blood ; Ducks ; virology ; Hepatitis Virus, Duck ; Hepatitis, Viral, Animal ; prevention & control ; virology ; Neutralization Tests ; Picornaviridae Infections ; prevention & control ; veterinary ; Poultry Diseases ; prevention & control ; virology ; Vaccines, Inactivated ; immunology ; Viral Hepatitis Vaccines ; immunology

To reveal the genetic variation of the viral protein 1 (VP1) gene of the duck hepatitis A virus type 3 (DHAV-3), the VP1 gene of 13 virulent DHAV-3 strains isolated from Shandong province of China in 2012 were amplified by RT-PCR, sequenced and analyzed. The results showed that all the VP1 genes of the 13 isolates contained 720 nucleotides encoding 240 amino acids, and shared with nucleotide identities of 94. 6%-99.9% and amino acid identities of 95.0%-100%. The nucleotide and amino acid sequence homologies between the 13 DHAV-3 isolates and other 31 DHAV-3 reference strains were 92.5%-100% and 90. 8%-100%, respectively. Phylogenetic analysis showed that the VP1 gene of DHAV-3 had distinct geographical characteristics. Distribution of genotypes of the 44 DHAV-3 strains was as follows: except the vaccine strain B63, all the other Chinese isolates belonged to genotype I (GI), Vietnamese wild isolates mainly belonged to subtype 1 (S1) of genotype II (GII), and all Korean isolates belonged to subtype 2 (S2) of GII.
Amino Acid Sequence ; Animals ; Capsid Proteins ; chemistry ; genetics ; China ; Ducks ; Hepatitis Virus, Duck ; classification ; genetics ; isolation & purification ; Hepatitis, Viral, Animal ; virology ; Molecular Sequence Data ; Phylogeny ; Picornaviridae Infections ; veterinary ; virology ; Poultry Diseases ; virology

OBJECTIVE: To study the expression of interferon-λ1 (IFN-λ1) in respiratory epithelial cells in children with human rhinovirus (HRV) infection. METHODS: Sputum samples and nasopharyngeal swabs were collected from the children who were hospitalized due to acute respiratory infection from February to October, 2017. Bacterial culture was performed, and nucleic acid test was performed for 11 respiratory pathogens. A total of 90 children with positive HRV alone were enrolled as the HRV infection group, and 95 children with positive respiratory syncytial virus (RSV) alone were enrolled as the RSV infection group. A total of 50 healthy children who underwent outpatient physical examination during the same period of time and had negative results for all pathogen tests were enrolled as the healthy control group. Nasopharyngeal swabs were collected from all groups, and quantitative real-time PCR was used to measure viral load and the mRNA expression of IFN-λ1. RESULTS: In the HRV infection group, there was no significant difference in the mRNA expression of IFN-λ1 between boys and girls and across all age groups (P>0.05). In the HRV infection group, there was no correlation between the mRNA expression of IFN-λ1 and HRV load (P>0.05). The mRNA expression of IFN-λ1 in the HRV infection group was significantly higher than that in the healthy control group (P<0.05), but significantly lower than that in the RSV infection group (P<0.05). CONCLUSIONS HRV can induce the expression of IFN-λ1 in respiratory epithelial cells, suggesting that IFN-λ1 may play an important role in anti-HRV infection in children.
Antiviral Agents ; Child ; Epithelial Cells ; Female ; Humans ; Interferons ; Male ; Picornaviridae Infections ; Respiratory Tract Infections ; Rhinovirus

In the present work, we reviewed the discovery, epidemiology, molecular biology and detection of Kobuvirus. Future fields of research were also discussed.
Animals ; Genome, Viral ; Humans ; Kobuvirus ; genetics ; isolation & purification ; Picornaviridae Infections ; epidemiology ; veterinary ; virology

OBJECTIVEWe want to explore the harm degree of human rhinovirus in infants in Beijing area.

METHODSFrom May 2008 to September 2009, 240 nasopharyngeal aspirates were collected from the children and infants who were hospitalized and with lower respiratory tract infections. These specimens were screened for HRV by real-time reverse transcription PCR (RT-PCR) and statistically analysised.

RESULTIn all of 240 hospitalized children, 208 cases were admission diagnosis of pneumonia, accounting for 86.67% (208/240), no deaths, the ratio of male and female patients was 1.93 : 1, and the collected samples reached to a maximum number in February 2009. Real-time PCR used to detect human rhinovirus, positive samples number is 71, positive rate is 29.58% (71/240), and the main symptoms and clinical diagnosis was pneumonia. Most cases were less than 2 years old, making up 81.69% (58/71), amony them, 13 months-18 months age and > or = 24 months groups have the highest incidence rates, the incidence rate is 33.33%.

CONCLUSIONHuman rhinovirus happened in spring and winter seasons, especially the infants who were under 2 years are the main infection groups, the important symptoms are lower respiratory infections such as pneumonia, bronchitis and bronchiolitis et al. Human rhinovirus is seasonal and contagious, spreads fast, so protective measures in hospitals should be prepared to avoid cross-infection.

Child ; Child, Preschool ; China ; Female ; Humans ; Infant ; Male ; Picornaviridae Infections ; virology ; Respiratory Tract Infections ; virology ; Rhinovirus ; genetics ; isolation & purification ; Seasons