Genetic characterization of echovirus 6 isolated from meningitis and encephalitis cases in Shandong Province, China / 病毒学报

To analyze the genetic characteristics of echovirus 6 (E6) isolated from meningitis and encephalitis cases in Shandong Province, China, we collected cerebrospinal fluid samples from meningitis and encephalitis cases in Shandong Province from 2007 to 2012 for virus isolation. Viral RNAs were extracted from positive isolates, and complete VP1 coding regions were amplified by RT-PCR and sequenced. Homology comparison and phylogenetic analysis were performed. Six isolates were identified as E6 by microneutralization assay and molecular typing. The homology analysis showed that the six isolates had 78. 6%-99. 8% nucleotide and 95. 5%-100. 0% amino acid identities with each other, as well as 76. 9%-78. 4% nucleotide and 92. 3%-95. 1% amino acid identities with the prototype strain (D' Amori). The phylogenetic analysis based on the integrated VP1 sequences indicated that all Shandong E6 isolates could be separated into four clusters, designated as A, B, C, and D. The six E6 isolates belonged to clusters A, B, and D. Our study reveals high genetic differences between Shandong E6 isolates and suggests different transmission lineages of E6 co-circulated in Shandong Province.

Temporal fluctuation and genetic characterization of echovirus 11 isolates upon surveillance of environmental sewage in Shandong Province, China / 病毒学报

We wished to analyze the genetic characterization of echovirus 11 (Echo11) from samples of environmental sewage in Shandong Province (China). The VP1 coding region was typed as the strains were amplified. Phylogenetic analyses on the VP1 sequences from these isolates, strains isolated from AFP cases in the period 1994-2010 and others published in GenBank were conducted. From 2011 to 2012, 94 Echo11 strains were isolated from samples of environmental sewage in Jinan and Linyi City in Shandong Province. Numbers of Echo11 were seasonal and reached peaks in the summer and autumn in both cities; A- mong these isolates, nucleotide (nt) identities were 89.5%-100.0% whereas amino acid (aa) identities were 95.4%-100.0%. The nt and aa identities were 76.6%-79.7% and 90.4%-92.5% between those strains and the prototype (Gregory) strain of Echo11, respectively. All isolates from Shandong Province were the A genotype and the strains evolved very rapidly, which suggested that several transmission chains was co-circulating. We described the temporal fluctuation and genetic characterization of Echo11 isolates from surveillance of environmental sewage in Shandong Province, thereby providing important information for exploring the dynamic change and genetic variation of circulating human enteroviruses in this Province in China.

Seroprevalence of the newer enterovirus types A71, A90 and B87 in healthy individuals in Shandong Province, China / 病毒学报

This study aimed to investigate antibody levels of the newer human enteroviruses (EV) A71, A90, and B87 in the population of Shandong Province, and provide a scientific basis for the development of prevention and control measures. In this study, serum specimens were collected from 400 individuals living in Yantai city, Shandong Province in 2010. EV-A71, A90, and B87 antibodies were detected using neutralization tests, and the results were analyzed by statistical methods. It was found that the positive neutralizing antibody rates of EV-A71, A90 and B87 in the population were 46.0%, 8.8%, and 47.0%, respectively. Their geometric mean titers (GMT) were 1 : 5.20, 1 : 1.49, and 1 : 4.02, respectively. Positive antibody rates for EV-A71 and EV-B87 were lowest in the 1-yr and 7-mo age groups, respectively. Positive rates increased gradually with age, and become consistent in the population aged >5 years. Positive antibody rates of EV-A90 were consistent across all age groups. Maternal antibody levels of EV-A71 declined rapidly after birth, and the increase in seroprevalence among 3-7 years old children implied that most EV-A71 infections occurred in preschool and early elementary school children. High positive antibody rates of EV-B87 in healthy individuals, especially children, implied that there may be an immune barrier within the general population. The population monitoring of EV-A90 should be strengthened, as its positive antibody rate is low.

Isolation and characterization of a human enterovirus 74 isolated in Shandong Province, China / 病毒学报

Human Enterovirus HEV 74 is a new member of species Human enterovirus B (HEV-B). To understand its evolution and restructuring characteristics, we report the complete genome sequence of a HEV74 strain 05293/SD/CHN/2005(abbreviated as 05293) isolated from an acute flaccid paralysis (AFP) case in Shangdong Province, China, 2005. Analysis of the complete genomic sequence of 05293 showed that its genome was collinear with that of previously described 2 HEV74 strains, except for insertions and deletions at the 5'NTR and the 3 NTR regions. The complete genome sequence of strain 05293 displayed 80. 8% nucleotide and 96% amino acid identity to the prototype strain USA/CA75-10213, and 80. 6% and 95. 9% to another isolated strain Rikaze-136. The P1, P2 and P3 coding regions of strain 05293 displayed 81. 5%, 80. 0%, 79. 7% nucleotide and 95. 9%, 96. 0%, 96.2% amino acid identity to the prototype strain USA/CA75-10213, and 81. 9%, 78. 8%, 79. 5% and 95. 9%, 96. 1%, 95. 7% to strain Rikaze-136, respectively. The phylogenetic tree and Simplot analysis on 05293 and HEV-B genome sequences were performed, and the result indicated frequent recombination within HEV-B.

Economic burden and related factors on inpatients with HBV-related diseases in Shandong province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the economic burden of patients with acute and chronic hepatitis B, cirrhosis and liver cancer caused by hepatitis B virus (HBV).</p><p><b>METHODS</b>Cluster sampling was used on cases consecutively collected during the study period. Questionnaire survey was conducted and information on the expenses during hospitalization was collected from the hospital records and through interviewing those patients.</p><p><b>RESULTS</b>Yearly costs related to patients with acute hepatitis B, severe hepatitis B, chronic hepatitis B, cirrhosis, hepatocellular carcinoma were 66.7, 138.1, 127.4, 151.7 and 377.2 thousand Yuan, respectively.</p><p><b>RESULTS</b>from multiple linear regression model showed that the type of medical insurance scheme, annual days of hospitalization, classifications of HBV-related diseases and personal income were major influencing factors on the cost.</p><p><b>CONCLUSION</b>HBV infection caused considerable burden to families and the society, indicating that HBV infection control programs would bring huge potential benefits. The reform of insurance scheme should be administrated to promote social fairness.</p>

Preliminary analysis on the prevalence and causes of breakthrough hepatitis B virus infection among children in Shandong province, China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To know the prevalence and probable causes of breakthrough hepatitis B virus (HBV) infection among children born after the introduction of universal infant hepatitis B vaccination in Shandong province, China.</p><p><b>METHODS</b>The subjects of this study were selected from the provincial hepatitis B serosurvey conducted in 2006, who were born between 1992 and 2005 (aged 1-15 years) and were confirmed to have completed three or more doses of hepatitis B vaccine. Finally 3527 subjects were involved in this study and were investigated using a unified question are. Blood samples were collected from them to detect hepatitis B surface antigen (HBsAg), antibody against HBsAg (Anti-HBs) and antibody against hepatitis B core antigen (Anti-HBc). The parents of children positive for HBsAg were followed up. Blood samples were collected from their parents to detect for HBsAg. The rate and correlative factors of breakthrough HBV infection were gotten by single-factor and multiple-factor analysis.</p><p><b>RESULTS</b>For the 3527 subjects, the overall prevalence rates of breakthrough HBV infection were 3.15% (111/3527), which decreased while birth year grew (χ(2)(Trend) = 44.83, P < 0.01) , the rate of subjects born in 1992 was the highest (9.9%, 16/161) , subjects born in 2000 was the least (0.8%, 2/258) , the rate of the self-report positive HBsAg status of mother, father and the other family members (15.22%, 7/46;34.09%, 15/44;17.65%, 6/34) were higher than the negative (2.99%, 104/3481, 2.76%, 96/3483, 3.01%, 105/3493) (χ(2) values were 22.28, 13.97, 23.68, respectively, all P values were < 0.01) , timely first dose of hepatitis B vaccine (5.37%, 41/763) was higher than the subjects that not in time (2.53%, 70/2764) (χ(2) = 15.596, P < 0.01) . The overall prevalence rates of breakthrough chronic HBV infection was 1.08% (38/3527), which decreased while birth year grew (χ(2)(Trend) = 9.96, P < 0.05) , the rate of subjects born in 1992 was the most (3.1%, 5/161) , subjects born in 1997 was the least (0.4%, 1/261) , the rate of the self-report positive HBsAg status of mother, father and the other family members (13.04%, 6/46;29.55%, 13/44;17.65%, 6/34) were higher than the negative (0.92%, 32/3481;0.72%, 25/3483;0.92%, 32/3493) (χ(2) values were 62.62, 338.80, 88.44, respectively, all P values were < 0.05) , timely first dose of hepatitis B vaccine (1.83%, 14/763) was lower than the subjects that not in time (0.87%, 24/2764) (χ(2) = 5.16, P = 0.02) . Multiple factors analysis showed that compared to the negative, the self-report positive HBsAg status of father, mother increased the risk of breakthrough HBV infection,OR (95%CI) values were 3.73 (1.09-12.75) and 26.76 (11.86-60.37) , respectively (all P values were < 0.05) , compared with eastern cities, the risk of western cities were the highest (OR (95%CI) = 6.00 (2.50-14.40) , P < 0.05) the risk of children born in 1992-2001 was higher than those born in 2002 ( (OR (95%CI) = 1.91 (1.10-3.32) , P < 0.05) . Compared to the negative, the self-report positive HBsAg status of father, mother and the other family members increased the risk of breakthrough chronic HBV infection,OR (95%CI) values were 7.51 (1.44-39.17) , 99.99 (34.29-291.62) , 8.94 (1.81-44.10) , respectively (all P values were < 0.05) , compared with eastern cities, the risk of western rural areas were the highest (OR (95%CI) = 12.51 (2.78-56.25) , P < 0.05) , sharing tooth brush with the others increased the risk (OR (95%CI) = 8.67 (1.14-66.14) , P < 0.05) . Among HBsAg-positive children, those with HBsAg positive mother and father accounted for 12/23 and 6/19, respectively.</p><p><b>CONCLUSION</b>The prevalence of breakthrough HBV infection and breakthrough chronic HBV infection among children was low in Shandong province. Mother to infant transmission might be the main reason for the infection while the role of the horizontal transmission within the family shouldn't be ignored.</p>

Identification and genetic characterization of coxsackieviruses A2, 6, 8 and 12 isolated in Shandong Province / 病毒学报

In previous study, molecular typing method was performed to identify human enteroviruses (HEVs) isolates collected from acute flaccid paralysis (AFP) cases from 1989 to 2011 and hand, foot and mouth disease (HFMD) patients, and 8 HEV-A serotypes were identified. In order to explore the genotypes and molecular evolution characteristics of HEV-A in Shandong province, viral RNA of the remaining isolates was extracted and entire VP1 coding region was amplified, sequenced and identified with HEV-A primers. Another 7 HEV-A Shandong isolates were obtained, and identified as Coxsackievirus A (CVA) 2, 6, 8 and 12 by molecular typing method. Homologous comparison showed that the nucleotide acid identities of Shandong strains ranged from 80.8% to 85.0% with prototype strains. Phylogenetic analysis based on VP1 sequences indicated that CVA8 and CVA12 strains were genetically related with domestic strains. However, CVA2 and CVA6 strains were distinct from both domestic and foreign strains. In addition, multiple transmission chains of CVA2 and CVA6 existed within Shandong province. So far, a total of 12 HEV-A serotypes were identified in Shandong province. This study enriched the distribution of serotypes and genetic evolution characteristics of HEV-A isolates in Shandong, and revealed different transmission chains of CVA2, 6, 8, 12 serotypes co-circulated in Shandong province or in China.

Identification and molecular characterization of human enteroviruses type 74, 80, and 87 isolated in Shandong Province, China / 病毒学报

An increasing number of new types of human enteroviruses (HEV) have been identified with the application of molecular typing method based on VP1 sequence analysis. In this study, the non-polio enteroviruses (NPEV) isolated from acute flaccid paralysis (AFP) cases in Shandong Province, China were typed via molecular typing method, and 1 EV74, 3 EV80 and 1 EV87 strains were identified. Homologous comparison revealed EV74, EV80 and EV87 Shandong strains had 81.4%, 76.4%-81.7%, and 80.3% VP1 identities with prototype strains. Phylogenetic analysis suggested a remote genetic distance with other strains. This is the first report of EV74 and EV87 in mainland China, and the low isolation rate in AFP surveillance suggested these three serotypes has not been the predominant viruses in China.

Analysis of the spatial and temporal characteristics of measles patients younger than 1 year old in Shandong province / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the spatial and temporal characteristics of measles patients younger than 1 year old in Shandong province.</p><p><b>METHODS</b>A total of 5309 cases of measles, whose patients were younger than 1 year old in Shandong province between year 1999 and 2008 were collected. The epidemic features of measles were described, and the annual infant incidence was calculated. Software ArcGIS9.3 was applied to draw the spatial map of the disease, and the software GeoDa0.95i-beta was adopted to analyze the spatial autocorrelation.</p><p><b>RESULTS</b>The incidence among infants younger than 1 year old reported in Shandong province rose from 23.45/100 000 (206 cases) in 1999 to 269.60/100 000 (2791 cases) in 2008.5309 cases covered all month-aged infants under 1 year old, except 12 months old. Most patients (3494 cases) aged between 6 - 9 months old; especially the infants around 8 months old, accounting for 20.7% (1100/5309). The epidemic peak was between March and May, accounting for 45.5% (2414/5309). The spatial and temporal distribution features showed an up and down temporal trend and an increase from east to west in spatial trend. The global Moran's I values of measles incidence among infants in Shandong province were 0.346, 0.150, 0.396, 0.213, 0.477, 0.354 and 0.331 in year 1999, 2001 - 2002, 2005 - 2008 (P < 0.01) and 0.076 in year 2004 (P < 0.05). The local spatial autocorrelation analysis showed that southwest and northwest districts of Shandong were highly clustered districts of measles.</p><p><b>CONCLUSION</b>In Shandong, the measles incidence among infants younger than 1 year old rose obviously; especially the infants aged between 6-9 months age. The epidemic peak was between March and May. A positive spatial correlation was found, the disease showed a distinct regional distribution feature, and a cluster district was found.</p>

Genetic characterization of poliovirus isolates from environmental sewage surveillance in Shandong, 2010 / 病毒学报

To investigate the genetic characteristics of poliovirus isolates from environmental sewage surveillance in Shandong province, we collected sewage samples in Jinan and Linyi City. Serotyping and VP1/ 3D sequencing were performed on polioviruses isolated from the concentrated sewage samples, and VP1 mutation and recombination were analyzed. Thirty-two of sewage samples were collected, and polioviruses were detected in 10 of the samples with a positive rate of 31.3%. Eighteen Sabin strains were isolated including three type 1, nine type 2, and six type 3 polioviruses, and the number of nucleotide substitutions in VP1 coding region varied from 0 to 4. Recombination was found in three Sabin 2 and four Sabin 3 polioviruses. Analysis of neurovirulence sites of VP1 revealed that one Sabin 1 vaccine strain had a nucleotide change of A to G at nt 2749, one Sabin 2 strain had a nucleotide change of A to G at nt 2908, three Sabin 2 strains had a nucleotide change of U to C at nt 2909, and all six Sabin 3 strains had a nucleotide change of C to U at nt 2493. Poliovirus vaccine strains could be isolated from environmental sewage with a high rate of gene recombination and back mutation of neuvirulence-associated sites. None of wild-type poliovirus or vaccine-derived poliovirus was detected.

Study on the genetic characterization and molecular epidemiology of echovirus 11 strains isolated in Shandong province, China / 中华流行病学杂志

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.

Identification and phylogenic analysis of Coxsackie-virus B5 during an outbreak of aseptic meningitis in Shandong / 中华流行病学杂志

Objective To identify the pathogen that caused an outbreak of aseptic meningitis in Shandong province in 2005. Phylogenic analysis was carried out on Coxsackie-virus B5 (CVB5) which was isolated during this outbreak. Methods 78 stool and 58 cerebrospinal fluids (CSF) specimens were collected from some inpatients during this outbreak. Virus isolation and reverse transcription-polymerase chain reaction (RT-PCR) was then performed. Phylo-genetic trees based on entire and partial VP1 sequences were constructed among CVB5 isolates and others published in GenBank. Results The isolation rates of stool and CSF specimens were 38.5% (30/78) and 48.3% (28/58) respectively. Among the results of serotype identification and molecular typing of 58 positive isolates, 54 were identified as CVB5, 2 as ECHO24, 1 as CVB3 and 1 as CVA9. Results from viral investigation showed that CVB5 was the main pathogen causing this outbreak. Data from homological comparisons indicated that Shandong strains had the highest nucleotide acid identity with the Zhejiang/ 12/02 strain (97.5%-97.8%), and lower identity (78.3%-78.6%) with the prototype strain (Faulkner strain). Phylogenic tree in VP1 region showed that CVB5 could be separated into four genotypes. Isolates of this outbreak belonged to genotype D. Conclusion CVB5 was the major etiological agent correlated with this outbreak. The shift of predominant genotype might serve as one of the causes that associated with the outbreaks of aseptic meningitis.

Genotype distribution of enterovirus group B isolated in Shandong Province, China / 病毒学报

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.

Genotype distribution of enterovirus group C in Shandong Province, China / 病毒学报

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.

Analysis on molecular epidemiology of rubella virus in Shandong province during 2000-2007 / 病毒学报

Analyze the genetic characteristics of sixteen strains of wild-type rubella viruses derived from Vero cells, Rk13 cells or Vero/slam cells, and isolated from throat samples in Shandong province during 2000-2007. The 1107 nucleotide sequence of nucleoprotein (E1) gene of these isolates were amplified by RT-PCR, and the PCR products were directly sequenced. Comparing with the gene tree that was constructed based on the 739 gene sequences of the WHO reference strains, twelve isolated strains belonged to 1E genotype, one strain belonged to 1F genotype, three strains belonged to 2A genotype. The first strain belonged to 1E genotype was isolated in Shandong province in 2001, then genotype 1E became dominant genotype of wild rubella viruses circulated. The 1E genotype circulated from 2006-2007 was different compared with that circulated from 2001 to 2002, but no significant deviation in temporal and geographic distribution was found. The strain belonged to Genotype 1F was only isolated during 2000 to 2001. The three strains of 2A genotype of rubella viruses were similar to rubella viruses vaccine strain (BRDII). The most nucleotide mutation of rubella viruses among the sixteen strains were nonsense mutation, and the amino acid sequences were highly conservative with no change in important antigen sites. Alike the previous reports, there was the same amino acid mutation in protein E1 at the site of 338 in all of the 1E genotype rubella viruses isolated during 2001- 2007 in Shandong (Leu338 --> Phe338).

The analysis of "a" dominant mutation of hepatitis B virus in community-based population of Shandong Province, China / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To determine the rate and type of "a" dominant mutation of hepatitis B virus (HBV) in community-based population of Shandong province and the possible effect of hepatitis B vaccination upon "a" dominant mutation.</p><p><b>METHODS</b>The anticipants aged 1-59 years were selected by multi-stage random sampling from the general population of Shandong province. Hepatitis B vaccination status was obtained by inquisition (for those over 15 years old) or immunization record (for those under 14 years old). The blood samples were collected and detected for HBsAg by ELISA. HBV DNA was extracted from the sera with positive HBsAg and S gene was amplified by nested-PCR. The PCR produce was sequenced and compared with the standard sequence.</p><p><b>RESULTS</b>Overall, 7601 anticipants were investigated. HBV DNA was successfully amplified and sequenced in 102 of 239 samples with positive HBsAg. 14.70% sera samples mutated in HBV "a" determinant region and 13 mutation types were detected. There were no statistically differences in the mutation rate by age groups (born before or after national universal infant hepatitis B vaccination) and hepatitis B vaccination status.</p><p><b>CONCLUSION</b>The "a" determinant mutation seemed to be uncommon in community-based population of Shandong province and the mutation sites were relatively scattered. Hepatitis B vaccination has no effect on "a" dominant mutation of hepatitis B virus.</p>

Identification and genetic characterization of human enterovirus type 73, 75, and 97 strains of specie B isolated in Shandong province / 病毒学报

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.

Molecular characterization of coxsackievirus B3 isolated from an outbreak of aseptic meningitis in Shandong Province, China / 中华实验和临床病毒学杂志

<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>

Genotype distribution of enterovirus A species isolated in Shandong Province, China / 病毒学报

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.

Molecular epidemiological analysis of echovirus 19 isolated from an outbreak associated with hand, foot, and mouth disease (HFMD) in Shandong Province of China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To elucidate the genetic characterization and molecular epidemiological features of Echovirus 19 (E19) isolates collected from an outbreak associated with hand, foot and mouth disease (HFMD) in Tai'an city of Shandong Province of China from July to September, 2003.</p><p><b>METHODS</b>Thirty seven Echovirus 19 isolates were isolated from stool specimens and throat swabs collected during the outbreak, then major capsid (VP1) genomic sequence was determined, and phylogenetic tree was done based on the VP1 sequences among these 37 and other E19 viruses deposited in the Genbank. Also a representative strain named CHN-SD03-TN12 was selected for sequencing of 5'-untranslated regions (5'-UTR).</p><p><b>RESULTS</b>The identity rate was about 98.9%-100% among all these 37 E19 viruses. The genetic relationships between these 37 E19 isolates and other strains reported were also depicted. The identity rate was about 78.4%-78.9% compared with E19 reference strain Burke. The substitutions in the sequence of 5'-UTR resulted in changes in the conjectural properties of 5'-UTR of E19 viruses.</p><p><b>CONCLUSION</b>The genetic features of E19 viruses isolated during the outbreak in Shandong Province in 2003 may be associated with a genetic and antigenic drift that changes the virulence of the Shandong isolates, but the molecular changes in Shandong E19 viruses contributing to their phenotype remain to be further illuminated. However, the sequences described in this paper substantiate the changes taken place in capsid VP1 and 5'UTR regions. These substitutions may contribute to their tropism and virulence, and play a significant role in pathogenesis and clinical manifestations of the disease.</p>