Major changes in prevalence and genotypes of rotavirus diarrhea in Beijing, China after RV5 rotavirus vaccine introduction.

To analyze the epidemiological characteristics of group A rotavirus (RVA) diarrhea in Beijing between 2019 and 2022 and evaluate the effectiveness of the RV5 vaccine. Stool specimens were collected from patients with acute diarrhea, and RVA was detected and genotyped. The whole genome of RVA was sequenced by fragment amplification and Sanger sequencing. Phylogenetic trees were constructed using Bayesian and maximum likelihood methods. Descriptive epidemiological methods were used to analyze the characteristics of RVA diarrhea. Test-negative design was used to evaluate the vaccine effectiveness (VE) of the RV5. Compared with 2011-2018, RVA-positive rates in patients with acute diarrhea under 5 years of age and adults decreased significantly between 2019 and 2022, to 9.45% (249/634) and 3.66% (220/6016), respectively. The predominant genotype of RVA had changed from G9-VIP[8]-III between 2019 and 2021 to G8-VP[8]-III in 2022, and P[8] sequences from G8-VP[8]-III strains formed a new branch called P[8]-IIIb. The complete genotype of G8-VP[8]-III was G8-P[8]-I2-R2-C2-M2-A2-N2-T2-E2-H2. The VE of 3 doses of RV5 was 90.4% (95% CI: 28.8%-98.7%) against RVA diarrhea. The prevalence of RVA decreased in Beijing between 2019 and 2022, and the predominant genotype changed to G8P[8], which may be related to RV5 vaccination. Continuous surveillance is necessary to evaluate vaccine effectiveness and improve vaccine design.

Transmissibility quantification of norovirus outbreaks in 2016-2021 in Beijing, China.

The transmissibility is a crucial feature for norovirus, yet its quantitative estimation has been limited. Our objective was to estimate the basic reproduction number (R0 ) of norovirus and investigate its variation characteristics. Norovirus outbreaks reported from September 2016 to August 2021 in Beijing were analyzed. The susceptible-infected-removed compartment model was established to estimate R0 . Linear regression models and logistic regression models were used to explore the factors affecting the transmissibility of norovirus. The overall median R0 of norovirus was estimated as 2.1 (interquartile range [IQR] 1.8-2.5), with 650 norovirus outbreaks. The transmissibility of norovirus varied by year, outbreak setting and genotype. The R0 of norovirus during September 2019 to August 2020 (median 2.1, IQR 1.8-2.4) and September 2020 to August 2021 (median 2.0, IQR 1.7-2.3) was lower than that of September 2016 to August 2017 (median 2.3, IQR 1.8-2.7) (ß = 0.94, p = 0.05; ß = 0.93, p = 0.008). The R0 of norovirus for all other settings was lower than that for kindergarten (median 2.4, IQR 2.0-2.9) (primary school: median 2.0, IQR 1.7-2.4, ß = 0.94, p = 0.001; secondary school: median 1.7, IQR 1.5-2.0, ß = 0.87, p < 0.001; college: median 1.7, IQR 1.5-1.8, ß = 0.89, p = 0.03; other closed settings: median 1.8, IQR 1.5-2.0, ß = 0.90, p = 0.004). GⅡ.2[P16] outbreaks had a median R0 of 2.2 (IQR 1.8-2.7), which was higher than that for GⅡ.6[P7] outbreaks (median 1.8, IQR: 1.8-2.0, odds ratio = 0.19, p = 0.03; GⅡ.2[P16] as reference) and mixed-genotype outbreaks (median 1.7, IQR: 1.5-1.8, ß = 0.92, p = 0.02; mixed-genotype as reference). In kindergartens and primary schools, norovirus shows increased transmissibility, emphasizing the vulnerable population and high-risk settings. Furthermore, the transmissibility of norovirus may change over time and with virus evolution, necessitating additional research to uncover the underlying mechanisms.

Genotypic diversity and recombination of norovirus GI.6[P11] associated acute gastroenteritis outbreaks in Beijing, China, from 2016 to 2019.

Norovirus (NoV) is the leading pathogen responsible for global acute gastroenteritis (AGE) outbreaks and sporadic cases. NoV evolves through gene mutation and recombination, leading to the emergence of new strains capable of causing global epidemics. This study aimed to learn the epidemiological characteristics of 39 GI.6[P11] NoV outbreaks in Beijing, China, from 2016 to 2019 and to analyze the genetic diversity and phylogenetic process of GI.6[P11] strains. The Bayesian phylogenetic analysis of partial VP1 and RNA-dependent RNA polymerase (RdRp) genes showed that GI.6[P11] strains were clustered into four subclades. Eleven whole genome sequences were obtained through the amplicon sequencing with 16 pairs of newly designed primers. The phylogenetic trees based on the whole genome and ORF1, 2, and 3 showed that the clustering of the 11 strains was consistent with that of partial VP1 and RdRp genes. The Bayesian inference revealed that the most recent ancestor (TMRCA) for the four subclades of the phylogenetic tree based on the whole genome sequences was 2012.42, 2014.81, 2011.74, and 2015.53, respectively. The recombination sites of GI.6[P11] strains in Beijing were located near the ORF1/2 junction. The histo-blood group antigens (HBGA) binding sites of GI.6[P11] strains in Beijing were conserved and there were some unique amino acid mutations in non-structural proteins in the ORF1 region.

Genomic Analysis and Antimicrobial Resistance of Campylobacter jejuni Isolated from Diarrheal Patients - Beijing Municipality, China, 2019-2021.

Introduction: Campylobacter jejuni (C. jejuni) is the leading cause of human bacterial gastroenteritis worldwide and has a major impact on global public health. The objective of the present study was to conduct whole genome sequencing (WGS) to determine the genetic diversity, virulence factors, and determinants of antimicrobial resistance of C. jejuni during a 3-year surveillance period in Beijing, China. Methods: A total of 184 clinical isolates were obtained from sentinel hospital surveillance between 2019 and 2021. Antimicrobial susceptibility testing was conducted using the agar dilution method. WGS was employed to characterize the 184 C. jejuni strains. Results: Multilocus sequence typing analysis revealed high genetic diversity among the 184 C. jejuni strains, identifying 71 sequence types (STs) and 19 clonal complexes (CCs). The most prevalent ST was ST760 (6.5%), and the most common CC was CC21 (24.5%), consisting of 11 STs. High resistance rates were observed for ciprofloxacin (76.6%), nalidixic acid (76.1%), and tetracycline (71.2%). A total of 77 C. jejuni isolates (41.8%) exhibited multidrug resistance with 43 resistance patterns. Virulome analysis disclosed the differential distribution of virulence factors related to adherence, colonization, chemotaxis, as well as lipo-oligosaccharide and capsular polysaccharide biosynthesis. Resistome analysis demonstrated widespread resistance to quinolones and tetracycline, but low rates of macrolides resistance. The phylogeny, based on whole genome single nucleotide polymorphisms, indicated a high degree of clonality and grouped the C. jejuni strains into six clades. Closely related isolates that were part of a genetic cluster mostly shared a homogenous clonal complex. Conclusions: The present study emphasizes the rising resistance to quinolones and tetracycline, as well as the virulence potential and diverse genotypes identified among C. jejuni strains isolated from diarrheal patients in Beijing.

Spatio-temporal distribution and influencing factors of norovirus outbreaks in Beijing, China from 2016 to 2020.

BACKGROUND: Noroviruses are a leading cause of acute gastroenteritis (AGE) worldwide. The geographical characteristics of norovirus outbreaks in Beijing and their influencing factors remain unknown. This study aimed to explore the spatial distributions, geographical characteristics, and influencing factors of norovirus outbreaks in Beijing, China. METHODS: Epidemiological data and specimens were collected through the AGE outbreak surveillance system in all 16 districts of Beijing. Data on spatial distribution, geographical characteristics, and influencing factors of norovirus outbreaks were analyzed using descriptive statistics methods. We measured spatial, geographical clustering of high- or low-value deviance from random distribution using Z-scores and P-values as statistical significance measures with Global Moran's I statistics and Getis-Ord Gi in ArcGIS. Linear regression and correlation methods were used to explore influencing factors. RESULTS: Between September 2016 and August 2020, 1,193 norovirus outbreaks were laboratory-confirmed. The number of outbreaks varied seasonally, typically peaking in spring (March to May) or winter (October to December). Outbreaks primarily occurred around central districts at the town level, and spatial autocorrelation was evident in both the entire study period and in individual years. Hotspots of norovirus outbreaks in Beijing were primarily found in contiguous areas between three central districts (Chaoyang, Haidian, Fengtai) and four suburban districts (Changping, Daxing, Fangshan, Tongzhou). The average population numbers, mean number of all schools, and mean number of kindergartens and primary schools for towns in central districts and hotspot areas were higher than those in suburban districts and non-hotspot areas respectively. Additionally, population numbers and densities of kindergartens and primary schools were influencing factors at the town level. CONCLUSIONS: Hotspots of norovirus outbreaks in Beijing were in contiguous areas between central and suburban districts with high populations, and high kindergarten and primary school densities were the likely driving forces. Outbreak surveillance needs to focus on contiguous areas between central and suburban districts with increased monitoring, medical resources, and health education.

Asymptomatic Norovirus Infection Among Children in Kindergartens and Primary Schools - Beijing Municipality, China, 2021.

What is already known about this topic?: Children in kindergartens and primary schools are the high-incidence groups of norovirus acute gastroenteritis. However, asymptomatic norovirus infection among them is seldom reported. What is added by this report?: The norovirus positive rate was 3.48% among asymptomatic children in kindergartens and primary schools in Beijing Municipality in June 2021, the most common genotype was GII.4 Sydney, and no acute gastroenteritis outbreak was reported over the study period. What are the implications for public health practice?: The asymptomatic norovirus infection was relatively low among kindergarten children and primary school students in summer. Norovirus genotypes in asymptomatic children were similar to those circulating in the symptomatic cases. Asymptomatic norovirus infection may play a limited role in causing acute gastroenteritis outbreaks.

Prevalence and genetic diversity of Parechovirus A in children with diarrhea in Beijing, China, 2017-2019.

We analyzed the prevalence and genotypes of Parechovirus A (PeV-A) in children with diarrhea in Beijing, China, 2017-2019. A total of 1734 stool samples collected from children <5 years of age with diarrhea were tested for the presence of PeV-A. Viral RNA was detected by real-time RT-PCR, and then genotyped by nested RT-PCR. We detected PeV-A in 93 (5.4%, 93/1734) samples, of which 87 could be genotyped by amplification of either the complete or partial VP1 region or the VP3/VP1 junction region. The median age of PeV-A infected children was 10 months. Most PeV-A infections were observed between August and November, with a peak in September. Seven known genotypes of PeV-A1A, -A1B, -A3, -A4, -A6, -A8 and -A11 were detected and PeV-A1B was the most prevalent genotype. Coinfection with other diarrheal viruses was observed in 30.1% (28/93) of PeV-A positive samples. All strains of PeV-A1A, -A1B, -A4 and -A6 obtained in this study contained the arginine-glycine-aspartic acid (RGD) motif, while all strains of PeV-A3, -A8 and -A11 lacked it. This study revealed a high genetic diversity of PeV-A circulating in Beijing and PeV-A11 was reported for the first time in children with diarrhea in China.

Excess acute diarrhoea cases attributed to norovirus variants in Beijing, China between 2011 and 2018.

New norovirus (NoV) variants emerge often leading to increased acute gastroenteritis outbreaks and outpatient visits. However, these increases are rarely quantified. Between September 2011 and August 2018, we included a total of 133 131 acute diarrhoea cases in 11 enteric disease outpatient clinics which were open all year round in Beijing. Over the same period, the etiology surveillance for acute diarrhoea was conducted, a total of 13 139 specimens were collected and tested, and 16.84% (2213/13 139) of all specimens were NoV-positive. The partial VP1 genes were successfully sequenced in 965 NoV strains. GII.4 Sydney, GII.17, and GII.2 predominated in 2012-2013, 2014-2015, and 2016-2017, respectively. We estimated the excess NoV-associated acute diarrhoea cases using the adjusted Serfling regression model, and three excess periods were found, corresponding to the predominance periods of GII.4 Sydney, GII.17 and GII.2, respectively, represented increases of 180.5% (95% confidence interval [CI]: 115.0%-246.0%), 114.7% (95% CI: 66.4%-163.1%) and 152.6% (95% CI: 100.2%-205.0%), compared with the baseline level. New NoV variants often caused an excess in their first year of predominance, and the excess periods of NoV-associated acute diarrhoea cases coincided with the predominance periods of NoV variants.

Vibrio parahaemolyticus O10:K4: An Emergent Serotype with Pandemic Virulence Traits as Predominant Clone Detected by Whole-Genome Sequence Analysis - Beijing Municipality, China, 2021.

Introduction: Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen which causes gastroenteritis in humans, especially the O3:K6 pandemic clone which is still a prominent serotype in Beijing, China. In this study, we observed a novel serotype O10:K4 isolated from clinical diarrhea cases, which became the most prevalent clone in 2021. Methods: 73 clinical isolates were collected through sentinel hospitals' surveillance in 2021. Serum agglutination testing and antimicrobial susceptibility testing were conducted. Whole genome sequencing was applied to characterize 73 V. parahaemolyticus strains and complete phylogenetic analysis. Results: Seven serotypes were identified among 73 strains. O10:K4 was the most common serotype (83.6%), followed by O2:KUT, O4:KUT, and O1:KUT. Multilocus sequence typing divided the 73 isolates into 10 sequence types (STs) with ST3 as the most prevalent, which covered all O10:K4 strains. Most isolates were sensitive to common antimicrobial agents apart from colistin. All the O10:K4 isolates were positive for the thermostable direct hemolysin gene, toxRS/new, andorf8, and negative for the TDH-related hemolysin gene. The whole genome sequencing-single nucleotide polymorphism phylogenetic analysis revealed O10:K4 strains formed a main genetic lineage, which was genetically distinct from other serotypes. We also demonstrated the presence of two type III secretion system genes (T3SS1 and T3SS2) and ß lactamase resistance gene blaCARB-22 in all O10:K4 strains. Conclusions: The study confirmed the emergence of V. parahaemolyticus O10:K4 possessing virulence factors similar to the O3:K6 pandemic clone, which may have enabled them to become prevalent in Beijing, China.

Occurrence of respiratory viruses among outpatients with diarrhea in Beijing, China, 2019-2020.

Objectives: To investigate respiratory virus infections in diarrhea cases and identify the risk of respiratory virus transmission through feces. Methods: Fecal specimens were collected from diarrhea cases in enteric disease clinics in Beijing, China, from 2019 to 2020. Cases that tested negative for norovirus, rotavirus, sapovirus, astrovirus, and enteric adenovirus were included in the study. Real-time RT-PCR was used to detect 16 groups of respiratory viruses, and the major viruses were genotyped. Viruses isolation and digestion of clinical specimens and nucleic acid by artificial gastric acid or artificial bile/pancreatic juice were used to evaluate the risk of respiratory virus transmission through feces. Results: A total of 558 specimens were collected and 47 (8.42%) specimens were detected positive, 40 (13.33%, 40/300) in 2019, and 7 (2.71%, 7/258) in 2020, including 20 (3.58%) for human rhinovirus (HRV), 13 (2.32%) for Bocavirus (BoV), 6 (1.08%) for parainfluenza virus I (PIV), 4 (0.72%) for coronavirus (CoV) OC43, 3 (0.54%) for respiratory syncytial virus (RSV) A, and 1 (0.18%) for both BoV and CoV OC43. Syndrome coronavirus 2 (SARS-CoV-2) and other viruses were not detected in this study. Eight genotypes were identified in the 13 HRV specimens. BoVs 1 and 2 were identified in nine BoV specimens. HRV infectious virions were successfully isolated from 2 clinical specimens and clinical specimens of HRV, RSV, PIV, and CoV could not be detected after 4 h of digestion and their nucleic acid could not be detected after 2 h of digestion by artificial gastric acid or artificial bile/pancreatic juice. Conclusion: There may be a risk of respiratory virus transmission from diarrhea cases, and interventions against SARS-COV-2 epidemics are also effective for other respiratory viruses.

Epidemiological characteristics and genetic diversity of norovirus infections among outpatient children with diarrhea under 5 years of age in Beijing, China, 2011-2018.

BACKGROUND: Human noroviruses are the leading cause of sporadic cases and outbreaks of viral acute gastroenteritis in all age groups worldwide. METHODS: Epidemiological data and fecal specimens were collected between January 2011 and December 2018 from 4911 children < 5 years of age with diarrhea in three districts of Beijing. From 2011 to 2013, One-Step Reverse Transcription Polymerase Chain Reaction (RT-PCR) was used to detect noroviruses, and from January 2014 to December 2018, norovirus GI and GII were screened using duplex quantitative real-time RT-PCR (qRT-PCR). One-Step RT-PCR and RT-seminested PCR were performed to amplify the RNA-dependent polymerase and capsid genes of noroviruses in positive sample. Amplified products were sequenced directly; norovirus was typed using the online Norovirus Genotyping Tool v2.0 and phylogenetic analyses were conducted using MEGA-X. RESULTS: From 2011 to 2018, noroviruses were detected in 16.5% of specimens from children with diarrhea. The highest prevalence was observed in children aged 12 to 23 months (22.4%, 319/1421), followed by children aged 6 to 11 months (17.6%, 253/1441). The highest prevalence of norovirus infections occurred in autumn followed by winter, spring, and summer. From 2011 to 2018, the most prevalent dual types (genotype and polymerase type) were GII.4 Sydney[P31] (51.6%, 239/463), followed by GII.3[P12] (24.0%, 111/463), GII.4 2006b[P4 2006b] (7.3%, 34/463), GII.2[P16] (5.0%, 23/463), GII.17[P17] (2.6%, 12/463) and GII.6[P7] (2.6%, 12/463). GII.4 2006b[P4 2006b] predominated in 2011 and 2012. GII.4 Sydney[P31] predominated from 2013 to 2018. In total, 15 genotypes, 15 P-types and 19 dual types were detected in this study, reflecting the genetic diversity. CONCLUSIONS: There were significant epidemiological characteristics and genetic diversity among outpatient children with norovirus infections < 5 years of age in Beijing from 2011 to 2018. These characteristics differ from those of norovirus outbreaks in Beijing. The complete genome sequences of each genotype are needed to better understand norovirus evolutionary mechanisms.

Group A rotavirus prevalence and genotypes among adult outpatients with diarrhea in Beijing, China, 2011-2018.

Group A rotavirus (RVA) is one of the most common causes of severe diarrhea in children worldwide. However, RVA is also an important pathogen causing adult diarrhea, with higher infection rates in older patients. To provide evidence for rotavirus epidemic control and to inform vaccine development, we analyzed the molecular epidemiology of RVA among adult outpatients with diarrhea in Beijing from 2011 to 2018. Stool specimens were collected monthly from 14 districts. RVA was detected using enzyme-linked immunosorbent assay and real-time reverse-transcription polymerase chain reaction (RT-PCR). Genotyping of rotavirus was performed using multiplex semi-nested RT-PCR. Phylogenetic analysis was performed using maximum likelihood methods implemented in MEGA software (version 6.06). Logistic regression and chi-square tests were used to assess differences among age groups, districts, years, and genotype distributions. The prevalence of rotavirus was 10.16% (1310 of 12,893) among adult outpatients with diarrhea from 2011 to 2018 in Beijing. The highest prevalence (13.74%, 600 of 4367) was observed among those aged 41 to 65 years. November, December, and January had the highest positive detection rates. In 2011, G3P[8] and G9P[8] were the dominant genotypes. Starting from 2012, G9P[8] became the dominant genotype. Most G9 strains belonged to the G9-VI clade. Most P[8] strains belonged to the P[8]-III clade. RVA is a major cause of adult diarrhea in Beijing. Continuous molecular surveillance is needed, and transmission of rotavirus between children and adults should be investigated further.

Investigation of an imported cholera case in China with whole genome sequencing.

Determining the source and genetic characteristics of the imported pathogen is critical in the control of infectious diseases. Here, we reported the investigation of an imported cholera case in China in 2018 with a recent travel history in Nepal and India. Stool culture from the patient was identified as Vibrio cholerae serogroup O1, biotype El Tor, serotype Ogawa. The strain 2018HL24 possessed intact Vibrio seventh pandemic island I (VSP-I), Vibrio pathogenicity Island 1 and 2 (VPI-1, VPI-2). A VSP-II variant with a 13 kb deletion was also detected, which was identical to those observed in V. cholerae in cluster "Nepal-4". Phylogenetic analysis based on the core genome SNPs showed that the isolate was most closely related to the V. cholerae isolated in northern India not far from the border of Nepal in 2012 (16 SNPs). Combining the epidemiological data with phylogenetic analysis results, we speculate that the patient may got infected in Nepal-India region.

Molecular Characteristics of Klebsiella pneumoniae Isolates From Outpatients in Sentinel Hospitals, Beijing, China, 2010-2019.

Background:Klebsiella pneumoniae is an opportunistic pathogen associated with community-acquired and nosocomial infections. Since 2010, K. pneumoniae testing has been included into an existing diarrhea-syndrome surveillance system for estimating the prevalence of K. pneumoniae in diarrhea-syndrome patients, assessing antibiotic susceptibility, and investigating molecular characteristics of K. pneumoniae. Methods:Klebsiella pneumoniae strains were isolated from stool specimens from diarrhea-syndrome outpatients in Beijing, China. Isolates were tested for antibiotic susceptibility, and phylogenetic relationships were explored though whole genome sequence analysis. Multi-locus sequence type (MLST) alleles were extracted from the whole genome sequence (WGS) data. A maximum likelihood tree was generated by MEGAX. Genomes were annotated by Prokka; core genes were produced by Roary; a maximum likelihood phylogenetic tree was generated using FastTree. Results: Forty-four K. pnuemoniae strains were isolated from 2010 to July 2019; of these 37 were K. pneumoniae and seven were K. variicola. Antibiotic susceptibility testing showed that all 44 strains were sensitive to gentamicin, imipenem, amikacin, meropenem, kanamycin; 97.73% were sensitive to cefoxitin andlavo-ofloxacin; the highest antibiotic resistance rate was 79.55%, which was to ampicillin. We found three extended-spectrum beta-lactamase (ESBL) producing strains; we identified high-virulence ST types, including ST307 and ST65; and we found that ST23 has been the epidemic clone since 2010. MLST and core genome sequence analysis showed two distinct clusters of 44 K. pnuemoniae; 40 alleles were identified in core genome sequence analysis, while 36 alleles were identified in MLST typing. Conclusions: There is an urgent need for epidemiological and molecular studies to understand the dynamics of antibiotic resistance and virulence gene transmission to guide strategies for K. pneumoniae surveillance. WGS analysis provided high discrimination power and reliable and robust data useful for molecular epidemiology.

Norovirus outbreaks in Beijing, China, from 2014 to 2017.

OBJECTIVES: Noroviruses are a leading cause of acute gastroenteritis (AGE) outbreaks worldwide. This study examined the epidemiology and genetic characteristics of norovirus outbreaks in Beijing, China. METHODS: Epidemiological data and fecal specimens were collected through the AGE outbreak surveillance system in Beijing. GI and GII genogroup noroviruses were detected and genotyped. The data were analyzed using descriptive statistics. RESULTS: Between September 2014 and August 2017, 762 AGE outbreaks were reported in Beijing, of which 661 (86.7%) were laboratory-confirmed as norovirus. Most norovirus outbreaks were reported during the spring (66.9%, 442/661), occurred in kindergartens and elementary schools (92.3%, 610/661), and were caused by GII genogroup noroviruses (95.6%; 632/661). The genotypes of the norovirus strains were determined in 468 outbreaks, and GII.P16-GII.2 and GII.P17-GII.17 strains were the most commonly identified. GII.P17-GII.17 and GII.P16-GII.2 strains predominated in 2014-2015 and 2016-2017 outbreaks, respectively. GII.P16-GII.2 noroviruses were responsible for a steep increase in AGE outbreaks in Beijing: 549 norovirus outbreaks were reported from 2016 to 2017, 9.2 times the number that occurred during the previous year. CONCLUSIONS: Norovirus causes a large disease burden in Beijing, and the prevalence of non-GII.4 noroviruses presents a new challenge for the development of vaccines.

Prevalence and genotypes of group A rotavirus among outpatient children under five years old with diarrhea in Beijing, China, 2011-2016.

BACKGROUND: Rotavirus is a leading cause of severe diarrheal disease, and one of the common causes of death in children aged under five years old. The dominant epidemic strains may change in different years in the same area. In order to provide evidence for rotavirus epidemic control and inform vaccine development, we analyzed epidemiological patterns and genetic characteristics of rotavirus in Beijing during 2011-2016. METHODS: Stool specimens of outpatient children under five years old were collected from three children's hospitals on a weekly basis. Group A rotavirus antigens were detected using enzyme-linked immunosorbent assay (ELISA) kit. The partial VP4 genes and VP7 genes of rotavirus were both amplified and sequenced. Genotyping and phylogenetic analyses were performed. Logistic regression and Chi-square tests were performed to determine differences across age groups, districts and years in rotavirus prevalence and genotype distribution. RESULTS: A total of 3668 stool specimens from children with acute diarrhea identified through hospital-based surveillance were collected from 2011 to 2016 in Beijing. A total of 762 (20.8%) specimens tested positive for rotavirus. The rotavirus-positive rate was highest among the 1-2 years old age group (29.0%, 310/1070). November, December and January were the highest rotavirus-positive rate months each year. G9 was the most common G genotype (64.4%, 461/716), and P [8] was the most common P genotype (87.0%, 623/716) among the 716 rotavirus-positive specimens. G9P [8], G3P [8] and G2P [4] were the most common strains. The rotavirus-positive rates of samples in 2012 and 2013 were higher than that in 2011, and the dominant genotype changed from G3P [8] to G9P [8] in 2012 and 2013. VP7 gene sequences of G9 strains in this study clustered into two main lineages. Most of the G9 strains exhibited the highest nucleotide similarity (99.1%~ 100.0%) to the strain found in Japan (MI1128). VP4 gene sequences of P [8] strains were almost P[8]b. CONCLUSIONS: Rotavirus accounted for more than one fifth of childhood diarrhea in Beijing during the study period. Targeted measures such as immunization with effective rotavirus vaccines should be carried out to reduce the morbidity and mortality due to rotavirus.

Prevalence and antibiotic resistance of bacterial pathogens isolated from childhood diarrhea in Beijing, China (2010-2014).

BACKGROUND: Diarrhea is one of the main causes of morbidity and mortality among children less than 5 years of age worldwide, and its causes vary by region. This study aimed to determine the etiologic spectrum, prevalent characteristics and antimicrobial resistance patterns of common enteropathogenic bacteria from diarrheagenic children in Beijing, the capital of China. METHODS: Stool samples were collected from 2524 outpatients who were aged 0-5 years in Beijing, China during 2010-2014. Microbiological methods, real-time PCR and antimicrobial susceptibility test were used to identify the bacterial causes and antimicrobial resistance patterns in the isolates. RESULTS: Of the 2524 patients screened, we identified the causes of 269 cases (10.7 %) as follows: diarrheagenic Escherichia coli (4.6 %), Salmonella (4.3 %), Shigella (1.4 %) and Vibrio parahaemolyticus (0.4 %). Atypical EPEC, Salmonella enteritidis, Shigella sonnei and serotype O3:K6 were the most common serogroups or serotypes of the four etiological bacteria. The prevalence of pathogens was correlated with age, season and clinical symptoms. The highest proportion of all causative bacteria was found in children aged 3-5 years and in summer. The clinical symptoms associated with specific bacterial infection, such as fever; abdominal pain; vomiting; and watery, mucus, and bloody stool, were observed frequently in diarrheal patients. Salmonella showed moderate rates of resistance (40-60 %) to ampicillin, nalidixic acid, streptomycin and sulfisoxazole. Resistance to at least three antimicrobials was found in 50 % of isolates. Of the top three serotypes in Salmonella, high-level antimicrobial resistance to single and multiple antibiotics was more common among Salmonella typhimurium and Salmonella 1, 4, [5], 12:i:- than among S. enteritidis. More than 90 % of Shigella isolates showed more alarming resistance to most antibiotics, with a widened spectrum compared to Salmonella. CONCLUSION: Constant antibiotic surveillance is warranted because the bacteria were highly resistant to various antimicrobials. Our study contributes to the strengthening of the existing surveillance system and provides aid for effective prevention and control strategies for childhood diarrhea.

Increased norovirus activity was associated with a novel norovirus GII.17 variant in Beijing, China during winter 2014-2015.

BACKGROUND: Norovirus (NoV) is a leading cause of sporadic cases and outbreaks of acute gastroenteritis (AGE). Increased NoV activity was observed in Beijing, China during winter 2014-2015; therefore, we examined the epidemiological patterns and genetic characteristics of NoV in the sporadic cases and outbreaks. METHODS: The weekly number of infectious diarrhea cases reported by all hospitals in Beijing was analyzed through the China information system for disease control and prevention. Fecal specimens were collected from the outbreaks and outpatients with AGE, and GI and GII NoVs were detected using real time reverse transcription polymerase chain reaction. The partial capsid genes and RNA-dependent RNA polymerase (RdRp) genes of NoV were both amplified and sequenced, and genotyping and phylogenetic analyses were performed. RESULTS: Between December 2014 and March 2015, the number of infectious diarrhea cases in Beijing (10,626 cases) increased by 35.6% over that of the previous year (7835 cases), and the detection rate of NoV (29.8%, 191/640) among outpatients with AGE was significantly higher than in the previous year (12.9%, 79/613) (χ(2) = 53.252, P < 0.001). Between November 2014 and March 2015, 35 outbreaks of AGE were reported in Beijing, and NoVs were detected in 33 outbreaks, all of which belonged to the GII genogroup. NoVs were sequenced and genotyped in 22 outbreaks, among which 20 were caused by a novel GII.17 strain. Among outpatients with AGE, this novel GII.17 strain was first detected in an outpatient in August 2014, and it replaced GII.4 Sydney_2012 as the predominant variant between December 2014 and March 2015. A phylogenetic analysis of the capsid genes and RdRp genes revealed that this novel GII.17 strain was distinct from previously identified GII variants, and it was recently designated as GII.P17_GII.17. This variant was further clustered into two sub-groups, named GII.17_2012 and GII.17_2014. During winter 2014-2015, GII.17_2014 caused the majority of AGE outbreaks in China and Japan. CONCLUSIONS: During winter 2014-2015, a novel NoV GII.17 variant replaced the GII.4 variant Sydney 2012 as the predominant strain in Beijing, China and caused increased NoV activity.

Human calicivirus occurrence among outpatients with diarrhea in Beijing, China, between April 2011 and March 2013.

Human caliciviruses (HuCVs), including noroviruses (NoVs) and sapoviruses (SaVs), are the most common cause of diarrhea in adults and the second most common cause of diarrhea in children. Between April 2011 and March 2013, 3,832 fecal specimens were collected from outpatients with diarrhea from 17 hospitals in Beijing, China, and 669 specimens (17.5%) were positive for HuCV. Of the 287 HuCV-positive specimens, 263 (91.6%) were identified to be NoV, 23 (8.0%) were identified to be SaV, and one (0.3%) was identified to be a mixed infection of NoV and SaV. Of the 263 NoV-positive specimens, 237 (90.1%) were NoV GII, 21 (8.0%) were NoV GI, and 5 (1.9%) were a combination of NoV GI and GII. Among the 216 sequenced GII-positive samples, GII.4 was the most common genotype (70.4%, 152/216), followed by GII.13 (9.3%, 20/216). GII.4 Sydney_2012 was first detected in August 2012 and replaced GII.4 Den Haag_2006b as the predominant variant between September 2012 and March 2013. With the emergence of the GII.4 Sydney_2012 variant, 44.6% more patients with diarrhea visited the 17 hospitals (9,931 cases) than in the previous year (6,866 cases) between October and December 2012.