Objective To identify the pathogen that caused an outbreak of viral encephalitis in Henan area in 2011.Phylogenic analysis was carried out on Coxsackie-virus B5 (CVB5) which was isolated during this outbreak.Methods Five throat swab,21 stool and 14 cerebrospinal fluid (CSF) specimens were collected from 29 inpatients during this outbreak.Viral isolation and real time RT-PCR were then performed for all specimens.Viral nucleic acid of enterovirus 71 (EV71),coxsackievirus A 16 (CA16) and pan-enterovirus (PE)were detected by real time RT-PCR.Phylogenetic tree based on entire VP1 sequences was constructed among CVB5 isolates from 2 stool and 3 CSF specimens of 5 inpatients and others published data retrieved from GenBank.Results The real time RT-PCR results showed that the PE nucleic acid positive rates of throat swab,stool and CSF specimens were 60.0％ (3/5),61.9％ (13/21) and 85.7％ (12/14) respectively.All of these specimens were negative for EV71 and CA16.The isolation rates of throat swab,stool and CSF specimens were 20.0％ (1/5),25.0％ (5/21) and 29.0％ (4/14),respectively.BLAST with both VP1 and 5′-UTR sequences and molecular typing indicated that CVB5 was the main pathogen.Analysis among the 5 positve isolates based on the complete VP1 sequences showed 97.9％-99.5％ homology.Data from homologous comparisons indicated that these isolates had the highest nucleotide acid identity with the Changchun CVB5 CC10/10/Changchun strain (97.1％-98.1％) which caused hand,foot,and mouth disease (HFMD) outbreak in Changchun in 2010,and lower identity (89.0％-89.6％ and 91.8％-92.5％) with the COXB5/Henan/2010 and 03001N strain isolated from Pingdingshan,Henan in 2010 and 2012,respectively.Phylogenetic tree in VP1 region showed that isolates of this outbreak belonged to genotype D,the same clade with Changchun strain.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 this outbreaks.

China ; Electrophoresis, Gel, Pulsed-Field ; Escherichia coli O157 ; classification ; genetics ; Genes, Bacterial ; Humans ; Molecular Typing ; Virulence

Objective To investigate the antibiotic resistance and pulsed field gel electrophoresis (PFGE) patterns of clinical isolates of Salmonella (S.) typhi and S.paratyphi in Henan province during 2009-2011.Methods According to molecular typing and Salmonella K-B drug susceptibility test method published by international PulseNet bacterial infectious disease monitoring network and USA Clinical and Laboratory Standards Institute (CLSI),the drug susceptibility and PFGE molecule characteristics of 78 S.typhi and S.paratyphi strains isolated from sentinel hospitals in Henan were analyzed.Results The 78 strains orS.typhi and S.paratyphi were resistant to 13 kinds of antibiotics,in which 62 were multidrug resistant (79.5％),4 were resistant to 2-3 kinds of antibiotics (5.1％),41 were resistant to 5-8 kinds of antibiotics (52.6％),14 were resistant to 9-10 kinds of antibiotics (17.9％),3 were resistant to 11-12 kinds of antibiotics (3.8％).The resistant rate to cephalosporins,quinolones and other 3 kinds of antibiotic showed an increase trends.Seventy two strains ofS.typhi and S.paratyphi could be divided 14 molecular patterns by digestion with Xba Ⅰ and PFGE,each pattem contains 1-47 strains which shared the similarity of 66.03％-100.00％.Conclusions The drug resistance of clinical isolates of S.typhi and S.paratyphi was serious in Henan.The PFGE patterns showed diversity,but the predominant patterns could be still found.The PFGE patterns of some strains were associated with their drug resistance.

Objective: To investigate the antimicrobial resistance and pulsed field gel electrophoresis (PFGE) patterns of S.paratyphi A strains in Zhengzhou city isolated from sentinel hospitals in 2013-2015. Methods: According to Salmonella molecular typing and K-B drug susceptibility testing method published by international PulseNet bacterial infectious disease monitoring network and USA Clinical and Laboratory Standards Institute (CLSI2015), we analyzed drug sensitivity and PFGE molecular characteristics of 67 S.paratyphi A strains(11 strains in 2013, 7 strains in 2014, 49 strains in 2015) isolated from blood and stool samples in two sentinel hospitals of fever with rash syndrome surveillance system established in Zhengzhou city in 2013-2015. Results: The results showed 67 strains of S.paratyphi A had different levels of resistance to 13 kinds of antibiotics, 65 strains were multi-drug resistant strains (97.0%), 5 isolates were resistant to 2-3 kinds of antibiotics (7.5%), 41 isolates were resistant to 5-8 kinds of antibiotics (61.2%),11 isolates were resistant to 9-10 kinds of antibiotics(16.4%),8 isolates were resistant to 11-12 kinds of antibiotics(11.9%). 67 strains of S.paratyphi A were divided into 10 molecular patterns(PTYA1-PTYA10) by digestion with XbaⅠ restriction endonuclease and pulsed field gel electrophoresis, each pattern contains 1-48 strains with similarity ranged from 94.31%-100%. PTYA3 contained 48 strains, which was predominant band type; PTYA1, 9 contained 6 strains; PTYA 2, 4, 5, 6, 7, 8, 10 contained 1 strains among them. Conclusion The status of drug resistance of clinical isolates of S.paratyphi A in Zhengzhou city was rather serious, PFGE patterns showed diversity and dominant characteristics. The PFGE patterns of partial strains and its corresponding anti-drug spectrum have certain relevance and cluster relationship.

Objective: To investigate the infectious status, gene type transition and epidemiological features of rotavirus A isolated from infants and children (<59 months-of-age) in sentinel hospitals from 2008 to 2015 in Henan province, China. Methods: In total, 2 541 stool samples (each 3- 5 ml) were collected from infants and children aged less than five years in two sentinel hospitals and group A rotavirus was detected by a double antibody sandwich ELISA. Viral RNA was extracted from positive samples and G/P gene typing was performed using a two-step nested multiplex RT-PCR. Epidemiological information (including demographic information such as age, sex and clinical symptoms) was also collected from the patients and analyzed. Results: Group A rotavirus was detected in 30.9% (785/2 541) of diarrhea samples from children. The detection rate was higher in October (54.8%, 345/629) and lower in July (5%, 5/101) each year from 2008 to 2015. The group A rotavirus infection rate was higher in boys (30.6%, 451/1 476) than in girls (31.4%, 334/1 065) (χ²=0.18, P=0.664). Infection mainly occurred in 4-12 months old patients (61.3%, 481/785) (χ²=196.69, P<0.001), and the infection rate was lower in cities (26%, 258/992) compared with rural areas (34.0%, 527/1 549) (χ²=18.19, P<0.001). G typing of 785 strains of group A rotavirus revealed the following types: G1 (13.5%, 106 strains), G2 (11.1%, 87 strains), G3 (29.7%, 233 strains), and G9 (57.5%, 451 strains); P typing revealed the predominance of P[4] (11.3%, 89 strains) and P[8] (84.7%, 665 strains); gene type combinations comprised mainly G9P[8], G2P[4], G3P[8], G1P[8], respectively accounted for 52.9% (415), 9.7% (76), 17.3% (136), 11.3% (89). Gene type combinations G1 [8] and G3P[8] have been decreasing in prevalence since 2008 and G9P[8] has become the dominant gene type of group A rotavirus in Henan province. Among the group A rotavirus infection samples, the male:female infection ratio was 1.4∶1 (451/334), with no significant difference in the infection rate (χ²=0.18, P=0.664); the infection rate was higher in 4- 12 months old patients (61.3%, 481/785), with a significant difference detected between age groups (χ²=196.69, P<0.001). The rate of detection was lower in cities (26.0%, 258/992) than in rural areas (34.0%, 527/1 549) (χ²=18.19, P<0.001). Clinical analysis revealed a body temperature of below 37 degrees in 75.7% of positive cases (594 patients), 37.0- 37.5 degrees in 17.2% of cases (135 patients), 37.6-38.0 degrees in 2.0% of cases (16 patients), and above 38 degrees in 5.1% of cases (40 patients), with most cases showing no fever or a mild fever. The frequency of episodes of diarrhea among the patients was 0- 3 times (21.1%, 166 cases), 4- 6 times (65.6%, 515 cases), 7- 9 times (8.0%, 63 cases), or 10- 15 times (5.2%, 41 cases), mainly showing mild and moderate diarrhea. Vomiting also varied in frequency among the patients from no vomiting (86.9%, 682 cases), 1-2 times (11.8%, 92 cases), 3 times (6.0%, 47 cases), and more than 3 times (0.4%, 3 cases). The occurrence of dehydration varied from no dehydration (86.9%, 682 cases), mild dehydration of 1%- 5% (12.1%, 95 cases), to severe dehydration of ≥5% (1.0%, 8 cases). Conclusion A higher infection rate of group A rotavirus was detected in children younger than five years of age with acute diarrhea in sentinel hospitals in Henan province, including part-mixed infection cases. A predominance of cases was detected in the autumn, and secondly the spring of each year. Gene type G9P[8] was most frequently isolated. The majority of patients displayed no fever, vomiting or dehydration. The cases with clinical symptoms of fever, diarrhea, vomiting and dehydration often showed mild disease.

OBJECTIVETo investigate the antimicrobial resistance status and pulsed field gel electrophoresis (PFGE) patterns of Salmonella Enteritidis (S.Enteritidis) strains in Henan province.

METHODSS. Enteritidis strains were isolated from seven sentinel hospitals from March 2011 to December 2013. According to molecular typing and Salmonella (Kirby-Bauer, K-B) drug susceptibility testing method published by the international PulseNet bacterial infectious disease monitoring network and USA Clinical and Laboratory Standards Institute (CLSI), we analyzed drug sensitivity of 8 kinds antibiotics and PFGE molecule characteristics of 120 S. Enteritidis isolates from seven sentinel hospitals.

RESULTSAmong 120 strains of S. Enteritidis, 77 were isolated from male patients, 43 from female patients. A total of 78 strains S. Enteritidis were isolated from young children ranged from 0 to 5 years old (65.0%), including 57 strains isolated from 6 months to 2 years old (47.5%). The isolated time mainly centralized on May to October of the year, 11 strains isolated in March-April (9.2%), 48 were in May-July (40.0%),54 in August-October (45.0%), 7 in other months, with a typical summer seasonal characteristics. The resistance rate of 120 strains S. Enteritidis to ampicillin was 50.0% (n=60); to ceftazidime was 14.2% (n=17), to cefotaxime was 18.3% (n=22); to cefepime was 5.8% (n=7); to nalidixic acid was 61.7% (n=74); to ciprofloxacin was 8.3% (n=10), to norfloxacin was 5.8% (n=7); to gentamicin was 42.5% (n=51); to streptomycin was 21.7% (n=26); to chloramphenicol was 30.0% (n=36); resistance to methicillin benzyl ammonium was 11.7% (n=14), compound sulfamethoxazole resistance rate was 71.7% (n=86); the tetracycline resistant rate was 47.5% (n=57). All 120 strains of S. Enteritidis had different levels of resistance to 8 kinds of antibiotics, all strains were multidrug resistant strains, 28 isolates were resistant to 3-4 kinds of antibiotics (23.3%), 38 isolates were resistant to 5-6 kinds of antibiotics (31.7%), 39 isolates were resistant to 7-8 kinds of antibiotics (32.5%). All 120 strains of S. Enteritidis were divided into 44 molecular patterns by digestion with XbaI and pulsed field gel electrophoresis. each pattern contained 1-35 strains with similarity ranged from 54.3%-100%. EN14 and EN19 were the main PFGE types, including 35 and 29 strains respectively.

CONCLUSIONThe status of drug resistance of clinical isolates of Salmonella in Henan province was rather serious, PFGE patterns showed advantages and partial strain's corresponding resistant spectrum have certain relevance and the same aggregation relationship.

Anti-Bacterial Agents ; pharmacology ; Child, Preschool ; China ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Microbial Sensitivity Tests ; Molecular Typing ; Salmonella Infections ; microbiology ; Salmonella enteritidis ; classification ; drug effects

Objective To investigate the etiological,antimicrobial resistance status and pulsed field gel electrophoresis (PFGE) patterns of S.enteritidis strains in Henan Province.Methods Totally 82 strains of S.enteritidis strains were isolated from seven sentiel hospitals which were able to detect multiple pathogens from April,2013 to December,2015.According to Salmonella K-B drug susceptibility testing and molecular typing method published by the USA clinical and Laboratory Standards Institute (2015),the drug sensitivity to 7 kinds of antimicrobial agents and PFGE molecule characteristics of 82 S.enteritidis strains were tested.The PFGE patterns were analyzed by BioNumerics 6.0 software based on international PulseNet bacterial infectious disease monitoring network.Results Forty-seven strains of S.enteritidis were isolated from male patients,35 strains isolated from female patients.A total of 56 S.enteritidis strains were isolated from young children aged from 0 to 5 years old,including 29 strains isolated from 6 months to 2 years old.The strains were mainly isolated between May and November of the year,and 15 (18.3％) strains were isolated between March and April,27(32.9％) strains between May and July,34 (41.5％) strains between August and October,6 (7.3％) strains were isolated in the rest time of the year,with a typical seasonal characteristics of summer and autumn.Sxity-four (78.0％)strains of S.enteritidis were resistant to ampicillin;23 (28.0％) strains were resistant to ceftazidime;39 (47.6％) strains were resistant to cefotaxime:13 (15.9％) strains were resistant to cefepime;55 (67.1％) strains were resistant to nalidixic acid;24 (29.3％) strains were resistant to ciprofloxacin;32 (39.0 ％) strains were resistant to gentamicin;14 (17.1 ％) strains were resistant to chloramphenicol;47 (57.3％) strains were resistant to methicillin benzyl ammonium;13 (15.9％) strains were resistant to sulfamethoxazole and trimethoprim;21 (25.6 ％) strains were resistant to tetracycline.All strains weremuhi-drug resistant,and 21 (25.6％) isolates were resistant to 3-4 kinds of antibiotics,32 (39.0％)isolates were resistant to 5-6 kinds of antibiotics, 29 (35.4 ％) isolates were resistant to 7-8 kinds ofantibiotics.These 82 strains of S.enteritidis were divided into 25 molecular patterns by digestion withXba Ⅰ enzyme.Each pattern contained 1 to 26 strains with similarity ranged from 59.33％-100.00％.EN1 and EN2 were the main PFGE belt types and included 26 and 16 strains,respectively.Conclusions The drug resistance of clinical isolates of S.enteritidis in Henan province is serious.PFGE patterns show polymorphism and the dominant patterns,part of which are associated with drug-resistance spectrum and show aggregation.

Objective To detect and analyze the distribution of virulence factors,antimicrobial resistance and pulsed field gel electrophoresis (PFGE) patterns of Shigella sonnei,isolated in Henan province from 2011 to 2014.Methods Samples of diarrhea patients were collected and isolated with SS selective culture medium in 37 ℃ for 18-24 hours.All strains were identified under the Kligler iron agar/motility-indol-urea biochemical action and API20E biochemical system.Serological typing and prepared DNA template were carried out with thermal cracking method and multiplex PCR,to detect the virulence genes of Shigella sonnei.According to the molecular typing method and K-B drug susceptibility testing method published by the international PulseNet bacterial infectious disease monitoring network and USA clinical and Laboratory Standards Institute,antimicrobial susceptibility tests and PFGE molecular characteristics of these positive strains isolated from sentinel hospitals patients stool samples were analyzed.Results Among the 98 strains of Sonnei type Ⅰ and 118 strains of Sonnei type Ⅱ,all the strains carried carry different virulence genes including SHET-1B,SHET-2,ial,ipaH genes,with 4 kinds of virulence gene combination types.All the 216 strains of Shigella sonnei belonged to the multi-drug resistant strains,including 34 isolates resistant to 2-4 kinds of antibiotics (15.7％),147 isolates to 5-8 kinds of antibiotics (68.1％),24 to 9-10 kinds of antibiotics (11.1％),7 to 11 kinds of antibiotics (3.2％),and 4 to 13 kinds of antibiotics (1.9％).A total of 100 strains of Shigella sonnci were divided into 31 molecular patterns,digested by Xba Ⅰ and PFGE.Each pattern contained 1-13 strains with similarities ranged from 68.6％-100.0％.Conclusions All the Shigella sonnei strains carried virulence pathogenic factors,presenting serious status on drug resistance.PFGE fingerprinting patterns showed high polymorphism and dominant characteristics.PFGE patterns of partial strains and corresponding antidrug spectrum presented certain relevance and with same aggregation relationship.

Objective:To explore the serotype distribution and drug resistance of Shigella in stool samples of children under five years old with diarrhea from 2008 to 2017 in Sui County, Henan Province. Methods:A total of 4 721 stool samples of children under five years old with diarrhea were collected from Doufuyuan Clinic of Sui County during 2008 to 2017, and Shigella strains were isolated through bacterial culture. The slide agglutination test was used for serotyping of Shigella strains. Two hundred of seventy-one Shigella strains were selected in proportion, and multiple gradient polymerase chain reaction was used to detect virulence genes and Kirby-Bauer agar method was used for drug sensitivity. Trend chi square test was used to analyze the annual trend of drug resistance. Results:The detection rate of Shigella strains in 4 721 fecal samples was 20.69% (977/4 721). A total of 977 Shigella strains were divided into 13 serotypes in two groups, including 77.79%(760/977) Shigella flexneri strains and 22.21%(217/977) Shigella sonnei strains.The top three serotypes detected alternately every year.The dominant gene pattern of Shigella flexneri was Shigella enterotoxin ( shET)-1+ , shET-2+ , invasion plasmid antigen H ( ipaH)+ , invasion-associated locus ( ial)+ , accounted for 84.04%(179/213) and that of Shigella sonnei was shET-1-, shET-2+ , ipaH+ , ial+ , accounted for 46.55%(27/58). The drug resistance rates of 271 Shigella strains to ampicillin, nalidixic acid and tetracycline were more than 90% and the strains were more sensitive to cefepime and ceftazidime.The drug resistance rates to cefotaxime, cefepime, ciprofloxacin, chloramphenicol and sulfamethoxazole/trimethoprim increased year by year, and all had statistically significant differences ( χ2=24.027, 7.232, 6.039, 4.764 and 6.809, respectively, all P< 0.05). There were 98.52%(267/271) strains resistant to more than three kinds of drugs. The resistance rates of Shigella flexneri to ciprofloxacin, norfloxacin, and chloramphenicol were higher than those of Shigella sonnei, and the resistance rates to gentamicin and sulfamethoxazole/trimethoprim were lower than those of Shigella sonnei. The differences were statistically significant ( χ2=31.866, 14.868, 83.036, 68.534 and 14.738, respectively, all P<0.01). Conclusions:The major serotypes of Shigella in children under five years old in Sui County are constantly changing from 2008 to 2017. The dominant gene patterns of different serotypes are different. Most isolated strains have multiple drug resistances, and different serotypes have different resistance profiles.

Objective To investigate the drug tolerance and PFGE patterns of Salrnonella (S.) paratyphi A strains isolated from sentinel hospitals in Dengfeng,Henan province,during 2009-2015.Methods Venous blood samples were collected from paratyphoid patients and cultured in double phase blood culture bottle.Suspicious strains were identified and used for Salomonella.O antigen and H1/2 phase flagellum-induced serum agglutination test with API20E biochemical systems and SSI Salmonella typing sera.According to Salmonella molecular typing and K-B drug susceptibility testing method published by PulseNet China bacterial infectious disease monitoring network and USA Clinical and Laboratory Standards Institute,we analyzed the drug susceptibility and PFGE molecule characteristics of S.paratyphiA strains isolated from the patients.Results A total of126 strains of S.paratyphi A were isolated from 248 blood samples,the antigen modes of them were 1,2,12:a:-.The resistance rate of 126 strains of S.paratyphi A was 83.3％ to ampicillin;29.4％ to ceftazidime,31.2％ to cefotaxime,17.5％ to cefepime;62.6％ to nalidixic acid;19.3％ to ciprofloxacin,26.4％ to norfloxacin;22.8％ to gentamicin,47.9％ to streptomycin;19.2％ to chloramphenicol,24.2％ to methicillin benzyl ammonium,58.6％ to compound sulfamethoxazole and 46.7％ to tetracycline.The 126 strains of S.paratyphi A had different levels of resistance to 8 kinds of antibiotics,109 strains were multidrug resistant (86.5％),9 strains were resistant to 2-3 kinds of antibiotics (7.1％),76 strains were resistant to 5-8 kinds of antibiotics (60.3％),17 strains were resistant to 9-10 kinds of antibiotics (13.5％),7 strains were resistant to 11-12 kinds of antibiotics (5.6％).The 126 strains of S.paratyphi A were divided into 14 molecular patterns by digestion with Xba I and pulsed field gel electrophoresis.The antibiotics resistance to third generation cephalosporin (CAZ,CTX),one generation and three generation of quinolones (NAL,CIP,NOR) and aminoglycosides antibiotics (STR) showed an upward trend.Each pattem contained 1-98 strains with similarity ranged from 64.10％ to 100.00％.PTYA 1,6,9 and 10 were the main PFGE belt types.Conclusion The drug resistance of clinical isolates ofS.paratyphi A was serious in Dengfeng,Henan province,PFGE pattems showed a diversity,but predominant patterns could also be found.The PFGE patterns of some strains had clustering and were related with their antidrug spectrums.