OBJECTIVETo identification and analysis Aichi virus from diarrhea and normal children in Lanzhou, and discuss the relationship between Aichi virus and Infant Diarrhea.

METHODSAccording to the literature published data, Using RT-PCR method to amplified Aichi virus 3CD fragment and the positive products were sequenced and determined, and made the alignment analysis between the nucleotide sequences of the amplified fragment with the known sequence of this virus.

RESULTSThere was 1 case detection of Aichi virus in the 46 hospitalized children with diarrhea and 299 children with diarrhea out-patients specifically, Overall detection rate was 0.06%, and there was no Aichi virus was detected in normal control children. 2 viral 3CD gene and the known reference strains of nucleotide sequences were 97%, while phylogenetic analysis showed that genotype of 2 viral belongs to the B.

CONCLUSIONSThere existed B Genotype of Aichi virus in China, and more research is needed to clarified the etiology and epidemiology of Aichi virus characteristics.

Child ; China ; Diarrhea ; virology ; Feces ; virology ; Humans ; Infant ; Kobuvirus ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Picornaviridae Infections ; virology

Abstract: To investigate the distribution of avian influenza virus in environmental samples from live poultry markets (LPM) in China, samples were collected and tested by nucleic acid during 2009-2013 season. Each sample was tested by real-time RT PCR using flu A specific primers. If any real-time PCR was positive, the sample was inoculated into specific-pathogen-free (SPF) embryonated chicken eggs for viral isolation. The results indicated that the positive rate of nucleic acid in enviromental samples exhibited seasonality. The positive rate of nucleic acid was significantly higher in Winter and Spring. The positive rate of nucleic acid in LPM located in the south of China was higher than in northern China. Samples of Sewage for cleaning poultry and chopping board showed that higher positive rate of nucleic acid than other samples. The Subtype identification showed that H5 and H9 were main subtypes in the enviromental samples. Viral isolation indicated H5 subtypes was more than H9 subtypes between 2009 and 2013 while H9 subtypes increased in 2013. Our findings suggested the significance of public health based on LPM surveillance and provided the basis of prevention and early warning for avian flu infection human.
Animals ; China ; Feces ; virology ; Fresh Water ; virology ; Influenza A virus ; classification ; genetics ; isolation & purification ; Influenza in Birds ; virology ; Poultry ; Poultry Diseases ; virology ; Public Health ; Seasons ; Sewage ; virology

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.
Cell Line ; China ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; virology ; Feces ; virology ; Genotype ; Hand, Foot and Mouth Disease ; virology ; Humans ; Molecular Sequence Data ; Paraplegia ; virology ; Phylogeny

The current epidemic situation of coronavirus disease 2019 (COVID-19) still remained severe. As the National Clinical Research Center for Infectious Diseases, the First Affiliated Hospital of Zhejiang University School of Medicine is the primary medical care center for COVID-19 in Zhejiang province. Based on the present expert consensus carried out by National Health Commission and National Administration of Traditional Chinese Medicine, our team summarized and established an effective treatment strategy centered on "Four-Anti and Two-Balance" for clinical practice. The "Four-Anti and Two-Balance" strategy included antivirus, anti-shock, anti-hyoxemia, anti-secondary infection, and maintaining of water, electrolyte and acid base balance and microecological balance. Meanwhile, integrated multidisciplinary personalized treatment was recommended to improve therapeutic effect. The importance of early viralogical detection, dynamic monitoring of inflammatory indexes and chest radiograph was emphasized in clinical decision-making. Sputum was observed with the highest positive rate of RT-PCR results. Viral nucleic acids could be detected in 10%patients' blood samples at acute period and 50%of patients had positive RT-PCR results in their feces. We also isolated alive viral strains from feces, indicating potential infectiousness of feces.Dynamic cytokine detection was necessary to timely identifying cytokine storms and application of artificial liver blood purification system. The "Four-Anti and Two-Balance" strategy effectively increased cure rate and reduced mortality. Early antiviral treatment could alleviate disease severity and prevent illness progression, and we found lopinavir/ritonavir combined with abidol showed antiviral effects in COVID-19. Shock and hypoxemia were usually caused by cytokine storms. The artificial liver blood purification system could rapidly remove inflammatory mediators and block cytokine storm.Moreover, it also favored the balance of fluid, electrolyte and acid-base and thus improved treatment efficacy in critical illness. For cases of severe illness, early and also short period of moderate glucocorticoid was supported. Patients with oxygenation index below 200 mmHg should be transferred to intensive medical center. Conservative oxygen therapy was preferred and noninvasive ventilation was not recommended. Patients with mechanical ventilation should be strictly supervised with cluster ventilator-associated pneumonia prevention strategies. Antimicrobial prophylaxis was not recommended except for patients with long course of disease, repeated fever and elevated procalcitonin (PCT), meanwhile secondary fungal infection should be concerned.Some patients with COVID-19 showed intestinal microbial dysbiosis with decreased probiotics such as and , so nutritional and gastrointestinal function should be assessed for all patients.Nutritional support and application of prebiotics or probiotics were suggested to regulate the balance of intestinal microbiota and reduce the risk of secondary infection due to bacterial translocation. Anxiety and fear were common in patients with COVID-19. Therefore,we established dynamic assessment and warning for psychological crisis. We also integrated Chinese medicine in treatment to promote disease rehabilitation through classification methods of traditional Chinese medicine. We optimized nursing process for severe patients to promote their rehabilitation. It remained unclear about viral clearance pattern after the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Therefore, two weeks' quarantine for discharged patients was required and a regular following up was also needed.The Zhejiang experience and suggestions have been implemented in our center and achieved good results. However, since COVID-19 was a newly emerging disease, more work was warranted to improve strategies of prevention, diagnosis and treatment for COVID-19.
Betacoronavirus ; isolation & purification ; China ; epidemiology ; Coronavirus Infections ; diagnosis ; epidemiology ; therapy ; virology ; Disease Management ; Early Diagnosis ; Feces ; virology ; Humans ; Pandemics ; Pneumonia, Viral ; diagnosis ; epidemiology ; therapy ; virology ; Sputum ; virology

A simple, rapid and sensitive colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) method was established to detect norovirus genotype GII. The method employed a set of six specially designed primers that recognized eight distinct sequences of RNA-dependant RNA polymerase and capsid protein gene for amplification of nucleic acid under isothermal conditions at 65 degrees C for 60 minutes. The amplification process of RT-LAMP was monitored by the addition of HNB (Hydroxy naphthol blue) dye prior to amplification. A positive reaction was indicated by a color change from violet to sky blue and confirmed by agarose electrophoresis. The specificity of the RT-LAMP was validated by detecting several different diarrhea viruses including norovirus genotype GII. The sensitivity was determined by serial dilutions of RNA molecules from in vitro transcription of norovirus genotype GII in parallel with conventional RT-PCR detection. The assay was further evaluated with 93 clinical specimens of diarrhea patients. The results showed that the sensitivity of RT-LAMP was 1 000 copies/microL with a high specificity and the relative sensitivity was at the same level as that of conventional RT-PCR. Positive rate of RT-LAMP in analysis of clinical specimens was approximately the same as that of conventional RT-PCR as well. This colorimetric RT-LAMP assay was potential for rapid detection of norovirus genotype GII on spot due to the observation of visual result with high specificity and sensitivity, time-saving and cost benefit.
Caliciviridae Infections ; diagnosis ; virology ; Colorimetry ; methods ; Feces ; virology ; Genotype ; Humans ; Norovirus ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods

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.
China ; epidemiology ; Enterovirus B, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; epidemiology ; virology ; Feces ; virology ; Humans ; Molecular Sequence Data ; Phylogeny

Bronchoalveolar Lavage Fluid/virology* ; COVID-19/virology* ; Feces/virology* ; Genome, Viral ; Humans ; Mouth Mucosa/virology* ; Multiplex Polymerase Chain Reaction ; Nasal Mucosa/virology* ; SARS-CoV-2/genetics* ; Sputum/virology* ; Whole Genome Sequencing

OBJECTIVEEffects of RIDASCREEN Norovirus (C 1401) 3rd Generation kit (R-biopham AG, darmstadt, Germany) and IDEIA NLV kit (DAKOCytomation., Ely, UK) were compared for detecting human norovirus (HuNV) in fecal sample.

METHODSThe performance of the ELISA was compared with that of the reverse transcription polymerase chain reaction (RT-PCR) by testing a panel of 308 fecal samples collected from patients involved in outbreaks of gastroenteritis in Chang Chun and Guang Zhou. Gene sequencing was performed to positive samples tested by RT-PCR to determine genotype compared with standard sequences.

RESULTSRT-PCR is gold standard, RIDASCREEN Norovirus (C 1401) 3rd Generation kit had a high sensitivity of 96.10% but a specificity of 93.51%, and Dako kit had a low sensitivity of 95.83% but a high specificity of 95.76%.

CONCLUSIONRIDASCREEN Norovirus (C 1401) 3rd Generation kit is more Satisfactory for a preliminary screening.

Caliciviridae Infections ; diagnosis ; virology ; Disease Outbreaks ; Enzyme-Linked Immunosorbent Assay ; methods ; Feces ; chemistry ; virology ; Gastroenteritis ; diagnosis ; virology ; Humans ; Norovirus ; genetics ; immunology ; isolation & purification ; Reagent Kits, Diagnostic

OBJECTIVETo learn the epidemiologic characteristics of rotavirus diarrhea in three hospitals under sentinel surveillance from August 2001 to July 2004 and to provide background information for developing and implementing rotavirus vaccine.

METHODSData from hospital-based rotavirus surveillance among children < 5 years old hospitalized with acute diarrhea was used. Patients' clinic information and feces specimens were collected. Specimens were tested and typed for rotavirus.

RESULTSTotally, 3121 specimens were tested and the detection rate of rotavirus was 51%. Among all the children with rotavirus diarrhea, 94% were < 2 years old. G3 (69.9%) was the most prevalent serotype followed by G1 (6.6%) and G2 (2.9%). P[8] was the most common genotype of rotavirus. The most common G-P combination identified in this study was P[8] G3 (64.0%). Seveal other combinations of minor frequency were also identified.

CONCLUSIONRotavirus infection was most commonly seen among children < 5 years old hospitalized with acute diarrhea in the three hospitals. It is important to develop and implement rotavirus vaccine to prevent and control severe rotavirus infection. Because of the diversity of rotavirus strains, it is necessary to perform rotavirus strain surveillance to understand the dynamic nature of viral transmission.

Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Feces ; virology ; Female ; Hospitals ; statistics & numerical data ; Humans ; Infant ; Infant, Newborn ; Male ; Rotavirus Infections ; epidemiology ; virology ; Sentinel Surveillance

OBJECTIVETo investigate the epidemic of diarrhea caused by rotavirus infection among children treated at the Beijing Friendship Hospital in a period of 2 years.

METHODSStool samples were collected from 972 diarrheic children from January 2010 to January 2012. The antigen of rotavirus was detected by the gold imunochromatography assay (GICA).

RESULTAmong the samples studied, 370 out of 972 cases (38.1%) were positive for rotavirus antigen. The ratio of infected boys and girls was 2.14:1. Most infected children (91.4%) were under the age of two. The viral infection takes place all year round but breaks out mainly within a cold season that lasts from October through March, with a peak in November and December. Even in the spring and summer, however, its prevalence rates were quite significant among the children with diarrhea, ranging from 11.1% to 41.7%.

CONCLUSIONIn Beijing area, rotavirus is the major etiologic cause of diarrhea in children, especially in those under the age of 2. The peak prevalence occurs at any given time during the late fall and early spring seasons. It is necessary, therefore, to strengthen the surveillance of rotavirus infection in the spring and summer seasons in order not only to predict and prevent a potential outbreak in the end of the year, but also collect and provide more dynamic data for the further epidemiologic research of rotavirus diarrhea in the children.

Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Feces ; virology ; Female ; Humans ; Infant ; Male ; Prevalence ; Rotavirus ; genetics ; isolation & purification ; Rotavirus Infections ; epidemiology ; virology ; Seasons