Epidemiological characteristics and drug resistance surveillance of Shigella in infants and young children in Henan Province from 2013 to 2017 / 中华疾病控制杂志

Objective To investigate the pathogenic characteristics of Shigella in infants from 2013 to 2017 in Henan Province. Methods From 2013 to 2017, 606 Shigella strains were isolated from 5 149 children with diarrhea under 5 years old in Henan Province. Serotyping, drug sensitivity test and Polymerase Chain Reaction detection of virulence gene methods were used to detect the pathogen of Shigella. Results The detection rate of Shigella in children with diarrhea was 11.77%, and the highest detection rate was in the 1-2 age group(24.08%). 606 Shigella strains were divided into two groups and 11 serotypes. Shigella flexneri accounted for 73.43%， and Shigella sonnei accounted for 26.57%. Resistance of 176 Shigella strains to ampicillin and naphthidine was serious (resistance rate > 90%), and the resistance rates to chloramphenicol, ciprofloxacin, norfloxacin and compound sulfamethoxamine were higher than 65%, and the sensitivity of imipenem and cephalosporin were higher. There were differences in drug resistance between Shigella flexneri and Shigella sonnei. The virulence genes of infants were mainly shET-1+, shET-2+, ipaH+ and ial+, and 5 avirulent strains were detected. Conclusions The bacterial dysentery of infants in Henan Province is dominated by Shigella flexneri. There are serious resistance and multidrug resistance to common antibiotics, and the dominant genes in different serotyping strains are different.

Identification of Honeys and Syrups by Microwave Plasma Torch Mass Spectrometry / 分析化学

In this study, a method for rapid identification of three kinds of honey and four kinds of syrup by microwave plasma torch mass spectrometry ( MPT-MS) without sample pretreatment was established. Under the positive ion mode, honey and syrup were ionized by MPT-MS and analyzed by quadrupole mass spectrometry ( QMS). The MS data were analyzed with chemometrics methods. Consequently, principal component analysis ( PCA) showed that PC1, PC2 and PC3 had a total contribution rate of 91. 2％ , and cluster analysis (CA) indicated when the critical value was 7, this method could distinguish honey and syrup in addition to clover honey and inulin syrup. Partial least squares-discriminant analysis ( PLS-DA) also manifested that honey and syrup could be effectively distinguished. Discriminant analysis (DA) demonstrated that the discriminant accuracy of honey and syrup was 100％ . Accordingly, all the results showed that the method of MPT-MS combined with chemometrics could identify honey and syrup rapidly. In brief, this method has the advantages such as fast analysis speed, accurate information extraction, high recognition accuracy and without sample pretreatment, and can be used for the identification of honey and main adulterated syrup.

Hand-foot-mouth disease pathogen separation and EV71 VP1 gene analysis in Sanmenxia City, Henan Province, China / 病毒学报

The aim of this study was to understand the enterovirus types and biological features of pediatric cases of HFMD in Sanmenxia City during 2011, and compare the latter to a cohort of healthy children. Stool samples of 55 cases of HFMD and 60 healthy children were collected for the isolation and identification of enteroviruses using RNA extraction and real-time RT-PCR assays. EV71 and CA16 were identified by nucleotide sequencing using virus-specific VP1 primers; for the other enteroviruses, 012/011 and 008/013 primers were used for amplification and sequencing. The results were analysed by sequence alignment with known sequences, and the characteristics of the EV71 VP1 gene were also analyzed. The detection rates for enteroviruses in cases of HFMD and healthy children were 52.73% (29/55) and 18.33% (11/60), respectively. Among these, there were 22 cases of EV71, four cases of CA16 and three cases of other enteroviruses in the cases with HFMD. Eleven healthy children had intestinal viruses, of which nine were Coxsackie B virus strains (81.82%, 9/11). Gene sequencing of the 19 EV71 strains illustrated that they were all subgenotype C4a, but the evolutionary tree showed an obvious clustering between cases from Lingbao City and Lushi County. This study demonstrates that the EV71 subgenotype C4a and CA16 strains were the most common cause of HFMD in Sanmenxia City in 2011, and that Coxsackie B strains were prevalent in healthy children. This finding may indicate that there is a widespread source of recessive infection in the community.

Outbreak of dengue Fever in central China, 2013 / 生物医学与环境科学（英文）

In 2013, the first dengue fever (DF) outbreak in central China was reported in the central of Henan province, northern temperate regions, although they have been sequentially recorded in Southern China. 106 suspected DF cases were reported and 73 patients were confirmed dengue virus type 3 (DEN-3) infections. 62/392 (15.8%) local health persons showed DEN antibodies positive. To this day Henan is the northernmost province in China which has been reported about outbreak of DF and what is important is that it warns us the endemic range of DF has been expanded geographically in China.

Genome sequences of coxsackievirus B5 isolated from viral encephalitis patients in Henan province, 2011 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To analyze the complete genome sequences of two coxsackievirus B5 (CVB5) isolated in Henan province, 2011.</p><p><b>METHODS</b>Specimens were collected from viral encephalitis patients and followed by viral isolation on them. RNA were extracted from positive isolates and the amplified products were sequenced. The full-length genomes of them were acquired by assembling the fragments, using DNAStar 5.01 software while phylogenetic analysis were performed with Mega 5.05 and other software.</p><p><b>RESULTS</b>The genomes RNA of 03001N and 17Y showed 7408 bp and 7404 bp long, and the 5'- and 3'-untranslated regions were 747 bp, 743 bp and 103 bp, 103 bp, respectively. BLAST analysis of these two isolates, based on the complete genome, showed 97% identity, with both of them having the highest similarity(98%, 99%)to the CVB5 strain isolated from Henan in 2010 rather than other CVB5 strains. Coding regions of both isolates were 6558 bp, code for a polyprotein of 2185 amino acids (aa) and both of them showed 99% amino acid identity. Phylogenetic tree in VP1 region showed that the two isolates belonged to the same clade with other strains isolated from all over the country in the past years, except for some CVB5 strains isolated from Henan and Shandong province in 2009 that formed the other cluster.</p><p><b>CONCLUSION</b>It seemed that more than one group of CVB5 were circulating in Henan province and these two isolates appeared the main epidemic strains circulating in the past years.</p>

Comparison of different molecular assays for the rapid detection of enterovirus 71 (EV71) / 病毒学报

Molecular detection of enterovirus (EV)71 RNA based on PCR methods is a quick and sensitive approach. At present, different PCR-based methods for EV71 RNA detection are available, but comparisons of results obtained using different approaches are limited. This study is to compare the analytical sensitivity and specificity of different real-time reverse transcription-polymerase chain reaction (rRT-PCR) and conventional reverse transcription-polymerase chain reaction (cRT-PCR) assays for enterovirus and EV71 detection, Altogether, three rRT-PCR assays and one cRT-PCR assay targeting the 5'UTR gene for universal detection of enterovirus; two rRT-PCR assays andone cRT-PCR assay targeting the VP1 gene for specific detection of EV 71 were examined. All assays showed good specificity. The detection sensitivity ranged from 8.19 x 10 to 8.19 x 10(5) copy equivalents. In general, rRT-PCR assays were more sensitive than cRT-PCR assays. All rRT-PCR assays showed 100% sensitivity for clinical specimens.

Genetic characteristics of enterovirus 71 VP1 and epidemiology of hand-foot-mouth disease in Xinxiang in 2011 / 病毒学报

The study was performed to examine the enterovirus 71(EV71) VP1 genetic feature and the epidemiology of hand-foot-mouth disease (HFMD) in Xinxiang in 2011. Real-time RT-PCR was used for the detection of Pan-enterovirus, Coxsackievirus A 16(CA16) and EV71 from stool specimens of HFMD. The VP1 region was amplified from 10 EV71 positive samples and the products were sequenced. EV71 genotypes were characterized by homology and phylogenetic tree analyses. Additionally, epidemic data of Xinxiang HFMD in 2011 was analyzed. The results revealed that 73% of the specimens from severe cases were determined as EV71 positive, which was significantly higher than CA16-positive ones (19%) (P < 0.01). Ten EV71 strains isolated in Xinxiang belonged to C4a cluster of sub-genotype C4, with 2.8% nucleotide and 0.9% amino acid sequences divergence among them. At position 170 in VP1 gene, an alanine(A) was predominant in 9 isolates, while a valine(V) residue was observed in one isolate. Compared to the representative C4a strains which were closely related to Xinxiang isolates, the amino acid variations of the pre-dominant Xinxiang strains generally occurred at position 292, threonine --> alanine (T --> A). A total of 1118 HFMD cases were reported in Xinxiang in 2011, and 92% of them were younger than 3 years old; the incidence rate peaked in April and December, suggesting that it is very necessary to strengthen HFMD prevention and control even in cold weather.

Survey on ticks and detection of new bunyavirus in some vect in the endemic areas of fever, thrombocytopenia and leukopenia syndrome (FTLS) in Henan province / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the distribution, species, seasonal fluctuation of ticks and detect new bunyavirus in some hematophagus in the endemic areas of fever thrombocytopenia and leukopenia syndrome (FTLS) in Henan province.</p><p><b>METHODS</b>From March to December 2011, the free ticks were collected manually with white cloth from the grassland and the parasitic ticks were collected from the host skin by hand searching in Xinyang and Jiyuan. The density and seasonal fluctuation of ticks were analyzed after classification of the specimen. The hematophagus were collected including gadfly (38 in 16 groups), cattle lice (224 in 16 groups), mosquitoes (238 in 17 groups) and ticks (825 in 77 groups), then RNA of new bunyavirus were detected by RT-PCR.</p><p><b>RESULTS</b>A total of 12 388 ticks were collected in Xinyang and Jiyuan, consisting of 2 families, 5 geniuses and 6 species. In Xinyang city, 622 ticks were identified, consisting of 2 families, 3 geniuses and 3 species, including 2 (0.32%) Ornithodoros lahorensis, 451 (72.51%) Haemaphysalis longicornis and 117 (18.81%) Boophilus microplus. In Jiyuan city, 11 766 ticks were identified, consisting of 1 family, 4 geniuses and 5 species, including 7718 (65.60%) Haemaphysalis longicornis, 164 (1.39%) H.anatolicum anatolicum and 710 (6.03%) other ticks such as H. detritum, Boophilus microplus and Rhipicephalus sanguineus. Haemaphysalis longicornis were found in both districts as the predominant species in Henan province. Ticks were active from March to October. The average density was 160 per person hour and the peak was from May to July with density 278, 209 and 542 per person hour respectively. The results was positive in RNA detection of new bunyavirus in 11 groups of tick and 3 groups of gadfly by RT-PCR. The results were negative in all other hematophagus.</p><p><b>CONCLUSION</b>Ornithodoros lahorensis, Haemaphysalis longicornis, Boophilus microplus, H.anatolicum anatolicum, Rhipicephalus sanguineus and H. detritum were found in Henan province. Haemaphysalis longicornis was the predominant species. The density of ticks varied with the seasons. The detection of new bunyavirus by PCR was positive in some ticks and gadflies.</p>

Analysis of the epidemic characteristics of fever and thrombocytopenia syndrome in Henan province, 2007 - 2011 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze the epidemiological characteristics of fever thrombocytopenia and leukopenia syndrome (FTLS) in Henan province, China in 2007 - 2011.</p><p><b>METHODS</b>Data from specific surveillance system for FTLS in Henan and Information Management System of Chinese Center for Disease Control and Prevention were used to collect the information of the cases.Descriptive epidemiological methods were used to analyze the surveillance data during 2007 - 2011. Patients' sera were collected to detect new bunyavirus using fluorescent RT-PCR and virus isolation.</p><p><b>RESULTS</b>During 2007 - 2011, 1021 FTLS cases were reported in Henan province. The fatality rate was 2.25%with 23 deaths. The cases reported in Xinyang city were 1007, accounting for 98.75%.Cases were mainly occurred between April and October, accounting for 96.47% (985/1021). Epidemic peak was May to July, accounting for 59.16% (604/1021). The second peak occurred in September, accounting for 12.05% (123/1021). The age of the cases ranged from 1 to 88 years old with the median age of 59. Sex ratio (male:female) was 1:1.50 (408:613). In all cases, 93.73% (957/1021) were farmers. In 465 patients' sera, the positive rate of new bunyavirus was 69.25% (322/465) using fluorescent RT-PCR. In 164 patients' sera, 67 strains of new bunyavirus were isolated with isolation rate of 40.85% (67/164).</p><p><b>CONCLUSION</b>FTLS in Henan province is caused mainly by the new bunyavirus and has certain regional and seasonal characteristics. Most cases are female older farmers.</p>

Analysis on the diagnosis and treatment of a cluster of cases infected by new bunyavirus / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze and summarize the clinical characteristics, experience of diagnosis and treatment of cases infected by new bunyavirus, which occurred in Henan province in 2010.</p><p><b>METHODS</b>The clinical characteristics and effect of diagnosis and treatment of 5 cases were analyzed using descriptive epidemiological method. Blood specimens were detected by RT-PCR and pathogen separation.</p><p><b>RESULTS</b>PCR testing was positive for all 5 cases. New bunyavirus were isolated from 2 cases. In 5 cases, fever (5/5), the whole body aches (5/5), fatigue (5/5), anorexia (5/5), nausea (5/5), the chills (4/5), cough (4/5), expectoration (4/5), vomiting (3/5), conjunctival hyperemia (3/5); Leukocyte reduction (5/5), thrombocytopenia (5/5), elevated alanine aminotransferase (4/5), elevated aspartate aminotransferase (4/5), elevated lactate dehydrogenase (5/5), creatine kinase elevations (4/5), urinary protein (3/5). By symptomatic and supportive treatment and prophylactic antibiotics, the first case died and the other 4 cases were cured. The average course of disease was 15.4 days.</p><p><b>CONCLUSION</b>Cases infected by new bunyavirus have complicated clinical feature and multiple organ damage. If symptomatic treatment is in time, prognosis will be good.</p>

Establishment of indirect immunofluorescence assay (IFA) for detection of IgG antibody against new bunyavirus / 中华预防医学杂志

<p><b>OBJECTIVE</b>To develop an indirect immunofluorescence assay (IFA) for detection of IgG antibodies against new bunyavirus.</p><p><b>METHODS</b>The antigen slides were prepared with 5 new bunyavirus strains isolated using Africa green monkey kidney (Vero) cells. Specificity and sensitivity evaluation of IFA were carried out by optimizing working conditions of IFA. Using established IFA, serum samples from both acute and recovery phases were tested for 126 cases with fever thrombocytopenia and leukopenia syndrome in Xinyang, Henan province in 2007 - 2011. The results were compared with detections by RT-PCR.</p><p><b>RESULTS</b>The new bunyavirus stable immunofluorescence specific WZ69 strain was selected to prepare antigen slides of IFA. The optimum conditions of IFA were: optimum dilution for primary antibody (serum) and secondary antibody (isosulfocyanic acid fluorescence marked goat anti-human IgG antibody) was 1:40 and 1:150 respectively. The optimum dilution for Evans blue in secondary antibody was 1:20 000. Among the 126 patients, 96 paired serum specimens were tested positive to the new bunyavirus and 30 patients were tested negative to the virus. The positive rate of antibodies was 76.19%. There was no significant difference in results between IFA and RT-PCR (72.22% (91/126)) (P > 0.05).</p><p><b>CONCLUSION</b>The IFA has high sensitivity and specificity with easy operation. It can be used in detecting the new bunyavirus infection in patients with fever, thrombocytopenia and leukopenia syndrome.</p>

Culture, isolation and identification of new bunyavirus in African green monkey kidney(Vero) cells / 中华预防医学杂志

<p><b>OBJECTIVE</b>To culture, isolate and identify new bunyavirus in Vero cell line.</p><p><b>METHODS</b>Samples of 164 new bunyavirus positive by real time RT-PCR detection and well preserved serum specimens were selected from cases of fever, thrombocytopenia and leukopenia syndrome (FTLS) in Xinyang, Henan province in 2009 - 2011. These sera were cultured in Vero cell line and new bunyavirus were detected by observing cytopathic effect (CPE), Real-time RT-PCR, indirect immunofluorescence assay (IFA) and thin-section electron microscopy observation. A total of 10 positive PCR products were selected randomly for sequencing and the results were compared with sequence in Genbank.</p><p><b>RESULTS</b>Among 164 FTLS serum specimens cultured in Vero cell line, no special CPE were observed and 67 strains (40.85%) were positive detected by Real-time RT-PCR. Nucleic acid similarity of 10 specimens were 97.8% - 100% and there's also a high similarity (> 99%) between specimens and new bunyavirus isolates (Accession No. HQ141600.1). Among 67 positive strains, 58 of them showed specific fluorescence particles by IFA. The viral particles were observed to be spheres with a diameter of 80 - 100 nm by electron microscopy.</p><p><b>CONCLUSION</b>Vero cell line is suitable for culture, isolation and identification of new bunyavirus.</p>

Etiology surveillance of hand-foot-mouth disease in Henan province between 2008 and 2011 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To understand etiological types and distribution features of hand-foot-mouth disease (HFMD) in Henan province between 2008 and 2011.</p><p><b>METHODS</b>A total of 30 486 specimens of feces, rectal swabs or throat swabs from HFMD patients were collected by each Municipal CDC in Henan from 2008 to 2011. The enterovirus 71 (EV71), coxsackie virus A16 (CA16) and other enterovirus (EV) were detected by RT-PCR or real time RT-PCR. The VP1 gene of EV71 was amplified and the sequences were analyzed by bioinformatics software. A genetic evolution tree of the sequence was constructed as well.</p><p><b>RESULTS</b>The positive rates of EV71, CA16 and other EV were 62.70% (11 209/17 876), 12.03% (2150/17 876), 25.27% (4517/17 876) in 17 876 laboratory diagnosed cases, respectively. The differences were statistically significant (χ(2) = 157.17, P < 0.05). The positive rates of EV71, CA16 and other EV were 63.40% (7370/11 624), 11.58% (1346/11 624) and 25.02% (2908/11 624) in male patients and 61.40% (3839/6252), 12.86% (804/6252) and 25.74% (1609/6252) in female patients, respectively. The differences were statistically significant (χ(2) = 4.06, P < 0.05). The children under 5 years old were high-risk population of HFMD, accounting to 97.67% (17 459/17 876) of the laboratory-diagnosed patients.86.92% (15 537/17 876) cases were children between 1 to 3 years old. Constituent ratio of EV71 changed seasonally during a year, there was a high infection ratio of EV71 between April and June, especially in May, the infection ratio reached 69.34% (2384/3438). The positive rates of EV71, CA16 and other EV were 82.48% (5715/6929), 1.76% (122/6929) and 15.76% (1092/6929) among the 6929 laboratory-diagnosed severe cases, respectively. The positive rates of EV71 was higher than CA16 and other EV (χ(2) = 9259.17, 6170.81, P < 0.05, respectively). There were 117 deaths because of severe HFMD, 55 (47.01%) of which were laboratory confirmed. 50 death cases were infected by EV71, and according to the genetic evolution analysis, the VP1 gene of EV71 strain was belonged to subtype C4 of gene C.</p><p><b>CONCLUSION</b>The EV71 and CA16 were the main pathogens which caused HFMD in Henan province, and EV71 virus was the dominant strain, belonging to C4 subtype of gene C.</p>

Molecular epidemiological analysis of species B enteroviruses isolated from Henan Province of China during the six months in 2010 / 病毒学报

This report presents an overview of human enterovirus B species in Henan Province. A total of 14 isolates of HEV-B species isolated under HFMD surveillance network during the six months in 2010 were examined. Based on molecular typing results targeting VP1 region, 14 isolates were classified into 6 serotypes of HEV-B. Furthermore, comparison of these 14 isolates with reference strains and strains in mainland China was conducted. The phylogenetic analysis revealed that E25, E11 and E6 showed homology with those from Shandong Province which adjoins Henan Province. E1 and E13 showed homology with those from Yunnan Province, and E30 showed homology with Henan strain isolated in 2008. Cocirculation of two lineages of echovirus 6 was observed.

Genomic characteristics of coxsakievirus A16 isolated in Henan Province / 病毒学报

To reveal the genomic sequence characteristics of coxsackievirus A16 (CoxA16) strain isolated from patients with hand-foot-mouth disease (HFMD) in Henan province. A total of 406 samples were detected by reverse-transcription polymerase chain reaction (RT-PCR) and cell-culture-based isolation of coxsackievirus A16. The whole genome of CoxA16 isolate was amplified using 10 pairs of primers, the sequences were analyzed and phylogenetic tree was generated by bioinformatics software. The full length of HN1162/HN/CHN/2010 genome was 7411bp. Compared with the other CoxA16 strains released in GenBank, the nucleotide similarities were 87.0-97.9%, 77.0%-95.4%, 80.3%-96.9%, 77.9% 96.2%, 80.5-100% in 5'UTR, P1, P2, P3, 3'UTR region, respectively; The similarities of nucleotide and amino acid sequences in VP1 region were 91.4%-96.4% and 99.3%-99.7%, respectively. Phylogenetic tree analysis showed that CoxA16 strains isolated from Henan, Shenzhen, Guangzhou and Fujian belonged to the same cluster. The newly isolated CoxA16 from Henan province belonged to subgenotype C2/B-2. These results will have great significance in monitoring CoxA16 and for prevention and control of hand-foot-mouth disease.

Surveillance of healthy people's Japanese encephalitis neutralizing antibody in some areas of Henan Province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To analyze the prevalence characteristics and influence factors of Japanese encephalitis (JE) neutralizing antibody in healthy people.</p><p><b>METHODS</b>Xinyang and Luoyang is the two cities in Henan Province. In 2010, healthy people of these two cities were selected by random sampling method to eight age groups: less than one year old, 1 -2 years old, 3 -4, 5 -6, 7 -14, 15 -19, 20 -59,and above 60 years old, their blood specimens were collected in May before JE infection and in November after JE infection, then followed with epidemiological investigation for JE neutralizing antibody by MCPENT.</p><p><b>RESULTS</b>519 healthy people were surveyed, 1008 effective blood specimens were collected and tested. The JE neutralizing antibody positive rate was 59.52% in men, and 67.39% in male, these two rates had no statistical significance (chi2 = 3.41, P > 0.05). The JE neutralizing antibody was 58.66% in May, and 61.20% in November, these two rates had no statistical significance (chi2 = 0.68, P > 0.05). The JE neutralizing antibody positive rate of 0 - 14 years old age group was 55.19% in Xinyang, and 45.03% in Luoyang,these two rates had no statistical significance (chi2 = 3.53, P > 0. 05). The JE neutralizing antibody positive rate of above 15 years old age group is 97.78% in Xinyang,and 48.94% in Luoyang, these two rates had statistical significance (chi2 = 55.42, P < 0.05). The JE neutralizing antibody positive rate of JE vaccination was 56.85%, and 38.35% in no JE vaccination, these two rates had statistical significance (chi2 = 10.88, P < 0.05).</p><p><b>CONCLUSION</b>The JE neutralizing antibody positive rate was showing significant differences in people above 15 years old between Xinyang and Luoyang. The JE neutralizing antibody positive rate was showing significant differences between JE vaccination and no vaccination. Xinyang and Luoyang City, recommended strengthening the 0 - 14 year-olds immunized, and at the same time, exploring and paying attention to JE immunization strategy of people above 15 years old in Luoyang.</p>

Genomic characteristics and recombination of enterovirus 71 strains isolated in Henan Province between 2008 and 2010 / 病毒学报

To reveal the genetic features and recombination of enterovirus 71 isolates between 2008 and 2010. A total of 5 enterovirus 71 isolates were sequenced completely and phylogenetic analysis and recombination were performed. Phylogenetic analysis based on VP1 regions revealed that the Henan enterovirus 71 between 2008 and 2010 belonged to C4a in subgenotype C4. Bootscan analyses and phylogenetic analysis based on the 5'UTR, P1, P2, P3 genomic regions revealed the recombinations between EV71 genotypes B and C at the 2A-2B junction, and between EV71 genotype B and CA16 strain G-10 at the 3B-3C junction. Henan enterovirus 71 isolates between 2008 and 2010 belonged to C4a in subgenotype C4 which was the predominant virus genotype circulating in mainland China since 2004, a combination of intratypic and intertypic recombination were found in EV71 subgenotype C4.

Etiology survey on an outbreak of viral encephalitis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To isolate and identify the pathogen that caused an outbreak of viral encephalitis in Henan province in 2010; and to analyze the genetic characteristic of gene viral protein1(VP1) on the viral strains isolated.</p><p><b>METHODS</b>During the period of the outbreak of viral encephalitis in Lushan county, Pingdingshan city, Henan province, eight hospitalized patients were recruited in the study. All the patients' feces samples were collected. Three patients' cerebrospinal fluids samples and another four patients' serum samples were collected separately. The virus in the samples were isolated and identified by enterovirus (EV) combined serum. The VP1 gene of the positive isolate was amplified by reverse transcriptase PCR method, and its nucleotide sequence was detected and the genetic evolution was analyzed.</p><p><b>RESULTS</b>Fifteen samples were collected in total, including 8 feces samples, 3 cerebrospinal fluids samples and 4 serum samples. The results of Fluorescence Quota PCR detection showed that 11 out of 15 samples were positive; 2 strains of virus were isolated from 2 feces samples and the serotype were all Coxsackie-positive identified by the EV combined serum. The full-length VP1 genetic sequences were all 849 bp, and showed 77.1% - 96.9% similar to the nucleotide and 95.8% - 100% similar to the amino acid of CoxB5. The analysis showed that the genetic evolution tree was just the same with Genotype-D.</p><p><b>CONCLUSION</b>CoxB5 whose genotype was Genotype-D, was the pathogen that caused the outbreak of viral encephalitis in Lushan county, Pingdingshan city, Henan province.</p>

Genetic characterization of echovirus 25 isolated from Henan / 病毒学报

This research firstly reported the molecular analysis of ECHO25 (Entric Cytopathic Human Orphanviruses Type 25). To clarify molecular characteristics of the ECHO25 virus isolates in Henan Province and its relationship with the rest of world's isolates,the complete VP1 sequences of the 4 isolates in Henan were successfully amplified by RT-PCR and were compared with other ECHO25 isolates available from GenBank. Compared with the prototype strain JV-4, the nucleotide sequence identity was 79.2%-80.1%, and the amino sequence identity was 89.0%-92.4, the nucleotide sequence identity among the 4 strains isolates in Henan Province was 93.0%-99.0%, the amino sequence identity was 92.4%-97.5%. HN-01 and HN-26 strains had the highest level of homology, the nucleotide homology was 99.0%; All the 4 strains belonged to the B1 genotype.