ABSTRACT
Objective To analyze the evolution and genetic characterization of echovirus 11 (Echo11 ) from the acute flaccid paralysis (AFP) cases in Shandong province. Methods Isolation of Enterovirus was performed from stool samples of AFP cases from 1994 to 2009. All positive strains were sero-typed by neutralization test. Entire VP1 coding region from 27 strains typed as Echo 11 was amplified by reverse transcription-polymerase chain reaction(RT-PCR), and subsequently phylogenetic analyse on VP1 sequences from these strains and others published in GenBank were conducted. Results From 1994 to 2009, altogether 915 non-polio enterovirus (NPEV) strains were isolated with 79(8.6% ) isolates serotyped as Echo11. There were 876 nucleotides (nt) in the complete VP1 genes, encoding 292 amino acids (aa). The nt identities were 76.4%-100.0% among those Echo11 Shandong strains with the aa identities as 91.4% -100.0%. The nt and aa identities were 77.7%-80.7% and 90.7%-94.8% between Shandong strains and prototype strains, respectively.Conclusion All Echo11 strains could be divided into four genotypes. Shandong strains that forming three (A1, A2 and C1) new sub-genotypes, with every sub-genotype had several brands.Sub-genotype A1 appeared to be the lately circulating one.
ABSTRACT
Molecular typing was conducted for three human enteroviruses (HEV) isolated from acute flaccid paralysis (AFP) cases in Shandong province, China. RNAs from virus supernatants were extracted and complete VP1 genes were amplified by RT-PCR and sequenced. Genotypes of these isolates were identified as HEV type 73, 75 and 97, respectively by BLAST program. Homology and phylogenetic tree analyses were performed. Sequence analysis of VP1 gene showed significant variation compared with prototype strains. This study presents the genetic characteristics of HEV 73, 75 and 97 of specie B in Shandong Province, and the first report of HEV97 in China.
Subject(s)
Humans , Base Sequence , Cell Line , China , Enterovirus , Classification , Genetics , Enterovirus Infections , Virology , Molecular Sequence Data , Paralysis , Virology , Phylogeny , Viral Proteins , GeneticsABSTRACT
In order to explore the genotype distribution of human enterovirus group B (HEV-B) in Shandong Province and to study the correlation between HEV-B serotypes and disease outbreaks, we sequenced and analyzed the entire VP1 coding region of HEV-B isolated from acute flaccid parolysis (AFP) system and disease outbreaks in Shandong province during 1998-2008. All together twenty nine HEV-B serotypes were identified, including twenty Echovirus (ECHO) serotypes, five Coxsackievirus B (CVB1-5) serotypes, one Coxsackievirus A9(CVA9) serotype, and newer enteroviruses EV73, EV75, and EV97. E11, CVB3, E6, E14 and E25 were the five frequently isolated serotypes from AFP surveillance system. CVB3, CVB5 and ECHO30 were the major causative agent of aseptic meningitis in Shandong province. Comparison of nucleotide sequence homology showed 75.4%-99.6% inter-typic identities within Shandong strains, and 73.8%-85.2% identities with prototype strains. Amino acid sequence comparison showed the differentiation was not much. Our research showed different serotypes possessed distinct time-cycling pattern, and different sub-genotypes could be further classified according to the inter-typic genetic distance. Thereby the route and range of transmission of HEV could be determined.
Subject(s)
Humans , Cell Line , China , Enterovirus B, Human , Classification , Genetics , Genotype , Molecular Sequence Data , Phylogeny , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Human Enterovirus C group (HEV-C) includes 17 serotypes, which can not be serotype-identified by neutralization test using antiserum pool for NPEV. In order to elucidate the genotypes and molecular evolution of HEV-C in Shandong Province, We selected the strains isolated from AFP cases between 1994-2009 to perform reverse transcriptase-polymerase chain reactions (RT-PCR) by the primers specific for entire VP1 coding gene of HEV-C and sequencing. The phylogenetic tree was then constructed among these VP1 nucleotide sequences and other prototype strains. Totally 12 Shandong local strains were obtained and separated into 4 genotypes, CVA20, CVA21,CVA24 and EV 96. The homologous comparison and phylogenetic analysis showed Shandong strains were distinct from prototype strains in each genotype. This report showed that different genotype HEV-C strains spread widely in Shandong Province.
Subject(s)
Humans , Cell Line , China , Enterovirus C, Human , Classification , Genetics , Genotype , Molecular Sequence Data , Phylogeny , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
In order to study the genotypes and molecular evolution of human enterovirus (HEV) A species in Shandong Province, Stool samples were collected from AFP and HFMD patients in Shandong Province and virus isolation was performed. Reverse Transcription-Polymerase Chain Reactions (RT-PCR) specific for EV71 and CVA16 were performed with the virus isolates from HFMD patients. Positive isolates were selected for entire VP1 coding gene amplification and sequencing. Isolates with negative PCR results and isolates from AFP patients were selected for entire VP1 coding gene amplification and sequencing using primers specific for HEV A species. Phylogenetic tree was constructed among these VP1 nucleotide sequences and of other strains. Altogether 293 strains classified into 8 genotypes were isolated. The homologous comparison and phylogenetic analysis showed Shandong strains were distinct with prototype strains in every genotype. This report presents an overview of HEV-A in Shandong Province.
Subject(s)
Humans , Cell Line , China , Enterovirus A, Human , Classification , Genetics , Enterovirus Infections , Virology , Feces , Virology , Genotype , Hand, Foot and Mouth Disease , Virology , Molecular Sequence Data , Paraplegia , Virology , PhylogenyABSTRACT
<p><b>OBJECTIVE</b>To identify the pathogen caused an outbreak of aseptic meningitis in Tancheng county of Shandong province in 2008, and to analyze the molecular characterization of VP1 gene of the Coxsackievirus B3(CVB3) isolates.</p><p><b>METHODS</b>Stool and cerebrospinal fluid(CSF) specimens were collected from this outbreak for virus isolation with RD and Hep-2 cell. After typing by neutralization test, the VP1 gene of the isolates were amplified by RT-PCR and sequenced. Homologous comparison and phylogenetic analysis were performed.</p><p><b>RESULTS</b>35 strains of enteviruse were isolated from 22 stools and 120 CSFs(7 from stools and 28 from CSFs), 34 strains identified as CVB3 and 1 as Echovirus 30(ECHO30) by neutralization test. The nucleotide homologies were 90.5%-100.0% in the partial VP1 gene (381 bp) among 34 CVB3 isolates. Homology comparisons indicated that Shandong strains have the identity of 79.5%-81.6% with the CVB3 prototype strain Nancy. 012/2008TC/SD/CHN and 177/2008TC/SD/CHN showed the highest nucleotides homologies (98.2% and 91.0% respectively) with Fuyang19 strain of Anhui province in 2008 in complete VP1 gene. The phylogenetic tree based on complete VP1 genes showed that all the CVB3 correlated with aseptic meningitis in China recently came from the same evolution linkage and formed a monophyletic cluster.</p><p><b>CONCLUSION</b>The causative agent of this outbreak of aseptic meningitis was CVB3. CVB3 circulated in China was genetically different from other countries.</p>
Subject(s)
Female , Humans , Male , China , Epidemiology , Coxsackievirus Infections , Epidemiology , Virology , Disease Outbreaks , Enterovirus B, Human , Classification , Genetics , Feces , Virology , Meningitis, Aseptic , Epidemiology , Virology , Molecular Sequence Data , Phylogeny , Viral Proteins , GeneticsABSTRACT
Objective To comprehensively measure the burden of hepatitis B, liver cirrhosis and liver cancer in Shandong province, using disability-adjusted life years (DALYs) to estimate the disease burden attribute to hepatitis B virus (HBV)infection. Methods Based on the mortality data of hepatitis B, liver cirrhosis and liver cancer derived from the third National Sampling Retrospective Survey for Causes of Death during 2004 and 2005, the incidence data of hepatitis B and the prevalence and the disability weights of liver cancer gained from the Shandong Cancer Prevalence Sampling Survey in 2007, we calculated the years of life lost (YLLs), years lived with disability (YLDs) and DALYs of three diseases following the procedures developed for the global burden of disease (GBD) study to ensure the comparability. Results The total burden for hepatitis B, liver cirrhosis and liver cancer were 211 616 (39 377 YLLs and 172 239 YLDs), 16 783 (13 497 YLLs and 3286 YLDs) and 247 795 (240 236 YLLs and 7559 YLDs) DALYs in 2005 respectively, and men were 2.19, 2.36 and 3.16 times as that for women, respectively in Shandong province. The burden for hepatitis B was mainly because of disability (81.39%). However, most burden on liver cirrhosis and liver cancer were due to premature death (80.42% and 96.95%). The burden of each patient related to hepatitis B, liver cirrhosis and liver cancer were 4.8, 13.73 and 11.11 respectively. Conclusion Hepatitis B, liver cirrhosis and liver cancer caused considerable burden to the people living in Shandong province, indicating that the control of hepatitis B virus infection would bring huge potential benefits.