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1.
Chinese Journal of Emergency Medicine ; (12): 1521-1525, 2022.
Article in Chinese | WPRIM | ID: wpr-954575

ABSTRACT

Objective:To analyze the clinical characteristics and pathogenic distribution of severe pneumonia in adults in order to provide basis for clinical diagnosis and treatment.Methods:From June 2021 to April 2022, 145 patients with pneumonia admitted to the Department of Respiratory and Critical Care Medicine of the Second People's Hospital of Guangdong Province. According to whether they meet the diagnostic criteria for severe pneumonia, they were divided into severe ( n=63) and mild ( n=82) groups, and the clinical features between the two groups were compared. At the same time, the role of FilmArray detection in severe pneumonia was discussed. The measurement data were tested using independent sample t test or Mann-Whitney U test, and the counting data were tested using Chi-square test or Fisher exact probability method. Results:The age of the patients in the severe group was (72.67±1.71) years, male patients accounted for 84.1%, and the median hospitalization time was 16 days. Nine patients died in hospital; most of them had fever, shortness of breath, and change of consciousness, accompanied by hypertension, diabetes, cerebrovascular disease, chronic kidney disease, and tumor history. Compared with the mild group, the total number of leukocytes, neutrophil ratio, procalcitonin, and C-reactive protein were higher in the severe group, but the CD3 +, CD4 +, and CD8 + cell counts were lower ( P<0.05). The positive rate of FilmArray detection in the severe group was 81%, and the mixed infection of multiple bacteria accounted for 50%, which was higher than that of traditional culture ( P<0.05). The top four pathogens in severe group were Pseudomonas aeruginosa, Acinetobacter baumannii complex, Klebsiella pneumoniae, and Staphylococcus aureus, which were significantly higher than that in the mild group ( P<0.05). Resistance genes were detected in patients with severe disease, which was significantly higher than that in patients with mild disease (70.7% vs. 17.5%, P<0.05). Conclusions:Severe pneumonia is more common in elderly men, with more basic diseases and poor immunity. FilmArray has a high positive rate and can detect multiple pathogens, which may have a role in the rapid diagnosis of severe pneumonia.

2.
Chinese Journal of Experimental and Clinical Virology ; (6): 626-631, 2019.
Article in Chinese | WPRIM | ID: wpr-805389

ABSTRACT

Objective@#In this study, phage display technology was used to construct the human anti-Zika virus(ZIKV), phage antibody library and to obtain and express the monoclonal antibody. The aim was to master the preparation and expression of human phage antibody library screening method for highly specific antibodies.@*Methods@#The whole blood samples of Zika patients were collected and the lymphocytes were isolated. The RT-PCR method was used to amplify the antibody light chain and heavy chain Fab gene from lymphocyte Ig mRNA. The pComb3H system was used to construct the gene with genetic diversity Preparation of human anti-ZIKV phage antibody library. The purified antibody library was screened by using the purified ZIKV and the obtained ZIKV E protein antigen.@*Results@#The monoclonal antibody Fab fragment gene was successfully obtained for the ZIKV E protein antigen. The gene can be efficiently expressed in Escherichia coli.@*Conclusions@#According to the sequence analysis, this study showed that the monoclonal antibody was a new human genetically engineered antibody against ZIKV, which laid the foundation for the early diagnosis of ZIKV, and obtain a specific monoclonal antibody to ZIKV for human treatment of ZIKV infection.

3.
Chinese Journal of Microbiology and Immunology ; (12): 345-353, 2018.
Article in Chinese | WPRIM | ID: wpr-711411

ABSTRACT

Objective To investigate the infection status, serotype distribution, drug sensitivity and molecular characteristics of diarrheagenic Escherichia coli (DEC) in patients with diarrhea in Guangdong Province. Methods Fecal samples were collected, cultured and isolated by traditional methods. Suspected Escherichia coli isolates were confirmed by multiplex PCR used for detecting specific virulence genes and bio-chemical methods. Positive strains were serotyped, characterized for drug sensitivity and analyzed by pulsed-field gel electrophoresis ( PFGE). Results The total positive rate of DEC in patients with diarrhea was 6.26%. The positive rates of enteropathogenic Escherichia coli (EPEC), enterotoxigenic Escherichia coli (ETEC), enterohemorrhagic Escherichia coli (EHEC), enteroadherent Escherichia coli (EAEC) and en-teroinvasive Escherichia coli (EIEC) were 2. 47% , 1. 54% , 1. 32% , 0. 62% and 0. 09% , respectively, with infections primarily in children aged 0-<7 years. The total seropositive rate was 52. 54% , with EHEC accounting for 15. 00% . DEC showed high sensitivity to imipenem, ciprofloxacin, ceftazidime and cefo-taxime. The multidrug resistance rate of DEC was 58. 45% , with EPEC being the most serious for multidrug resistance. PFGE results showed that ETEC, EHEC, EPEC and EAEC had a high degree of polymorphism. Conclusion EPEC is the predominant type of DEC circulating in Guangdong Province. Third-generation cephalosporins are the first drugs of choice for treating infections in children. Ciprofloxacin can be used to treat adults. The problem of multiple drug resistance of DEC is severe and efforts to monitor DEC infections and drug resistance should be strengthened.

4.
Chinese Journal of Epidemiology ; (12): 63-66, 2018.
Article in Chinese | WPRIM | ID: wpr-737918

ABSTRACT

Objective To understand the circulation,drug resistance and molecular characteristics of Salmonella 1,4,[5],12:i:-in human in Guangdong province.Methods Salmonella 1,4,[5],12:i:-isolated from diarrhea patients in Guangdong during 2007-2016 were detected for drug resistance,genes and PFGE characteristics.Results A total of 2 960 strains Salmonella 1,4,[5],12:i:-were isolated from human diarrhea cases during this period.The positive rates of the isolation increased year by year.The male to female ratio of the infection cases was 1.58 ∶ 1,and the infection mainly occurred in infants and young children.Except imipenem,Salmonella 1,4,[5],12:i:-was resistant to other 17 antibiotics to some extent.The drug resistant rates to ceftazidime,cefotaxime and ciprofloxacin increased from 2011 to 2016.Multi-drug resistance was serious,for example,the multi-drug resistant strains with ASSuT accounted for 70.62% (435/616) and the multi-drug resistant strains with ACSuGSTTm accounted for 27.11% (167/616).The lack offljA,fljB and hin genes,as well as the retaining of iroB,STM2740,STM2757 genes,resulted in the unable expression of FljBenx gene with 8 different defection profiles.There were 934 different PFGE patterns observed in 2 347 strains,which displayed a relatively large fingerprint polymorphism.The major PFGE pattern was JPXX01.GD0226,which was found in 97 strains,accounting for 4.13% (97/2 347).The PFGE patterns in 168 Salmonella 1,4,[5],12:i:-strains were consistent with that of Salmonella typhimurium.Conclusions Salmonella 1,4,[5],12:i:-strains has become the major serotype of Salmonella that cause diarrhea in human in Guangdong.The multi-drug resistance of Salmonella 1,4,[5],12:i:-was serious,and since the defection offljA,fljB and hin genes,the expression of FljBenx protein failed.The PFGE results were diverse,which displayed polymorphism in inheritance.

5.
Chinese Journal of Epidemiology ; (12): 63-66, 2018.
Article in Chinese | WPRIM | ID: wpr-736450

ABSTRACT

Objective To understand the circulation,drug resistance and molecular characteristics of Salmonella 1,4,[5],12:i:-in human in Guangdong province.Methods Salmonella 1,4,[5],12:i:-isolated from diarrhea patients in Guangdong during 2007-2016 were detected for drug resistance,genes and PFGE characteristics.Results A total of 2 960 strains Salmonella 1,4,[5],12:i:-were isolated from human diarrhea cases during this period.The positive rates of the isolation increased year by year.The male to female ratio of the infection cases was 1.58 ∶ 1,and the infection mainly occurred in infants and young children.Except imipenem,Salmonella 1,4,[5],12:i:-was resistant to other 17 antibiotics to some extent.The drug resistant rates to ceftazidime,cefotaxime and ciprofloxacin increased from 2011 to 2016.Multi-drug resistance was serious,for example,the multi-drug resistant strains with ASSuT accounted for 70.62% (435/616) and the multi-drug resistant strains with ACSuGSTTm accounted for 27.11% (167/616).The lack offljA,fljB and hin genes,as well as the retaining of iroB,STM2740,STM2757 genes,resulted in the unable expression of FljBenx gene with 8 different defection profiles.There were 934 different PFGE patterns observed in 2 347 strains,which displayed a relatively large fingerprint polymorphism.The major PFGE pattern was JPXX01.GD0226,which was found in 97 strains,accounting for 4.13% (97/2 347).The PFGE patterns in 168 Salmonella 1,4,[5],12:i:-strains were consistent with that of Salmonella typhimurium.Conclusions Salmonella 1,4,[5],12:i:-strains has become the major serotype of Salmonella that cause diarrhea in human in Guangdong.The multi-drug resistance of Salmonella 1,4,[5],12:i:-was serious,and since the defection offljA,fljB and hin genes,the expression of FljBenx protein failed.The PFGE results were diverse,which displayed polymorphism in inheritance.

6.
Chinese Journal of Microbiology and Immunology ; (12): 611-617, 2017.
Article in Chinese | WPRIM | ID: wpr-613075

ABSTRACT

Objective To analyze the serotype distribution and antibiotic resistance characteristics of Salmonella strains isolated in Guangdong province for better understanding the condition of Salmonella infection in patients with diarrhea.Methods Fecal samples collected from patients with diarrhea in Guangdong province were used to isolate Salmonella strains.Biochemical analysis was performed to identify these isolated strains.Serotyping and antimicrobial susceptibility testing were carried out for further analysis of the isolated Salmonella strains.Results The rate of Salmonella infection was 7.64%in 2015, and the male to female patient ratio was 1.52∶1.A total of 2 377 patients of all age groups were positive for Salmonella infection and the patients aged 0-6 years accounted for 81.74%.The isolation rate of Salmonella strains in the summer and autumn was higher than that in the winter and spring (10.73% vs 4.24%;X2=463.77, P<0.01).The Salmonella isolation rates in different areas were as follows: 16.82% in Zhuhai, 15.85% in Heyuan, 11.81% in Yangjiang, 10.68% in Jiangmen, 8.49% in Zhongshan, 8.07% in Maoming, 8.05% in Jieyang, 7.35% in Shaoguan, 6.97% in Foshan, 6.03% in Dongguan, 5.48% in Guangzhou and 0.00% in Zhanjiang.And the differences between different regions were statistically significant (X2=367.67, P<0.01).The 2 377 isolated Salmonella strains were classified into 108 serotypes except for oneSalmonella strain that could not be classified.The top four predominant serotypes were 4,5,12:i:-, Salmonella enteritidis,Salmonella stanley and Salmonella typhimurium.Most Salmonella strains were sensitive to imipenem, azithromycin, ceftazidime, cefotaxime and trimethoprim/sulfamethoxazole, but multidrug resistance was common among those strains.Conclusion Salmonella serotypes of 4,5,12:i:-and Salmonella enteritidis are the predominant pathogens causing human Salmonella infections in Guangdong province.Ceftazidime and cefotaximeare are preferred in the treatment of Salmonella infections.Surveillance for drug resistance in Salmonella should be strengthened as multidrug resistant strains have become a serious problem in Guangdong province.

7.
Chinese Journal of Experimental and Clinical Virology ; (6): 409-413, 2017.
Article in Chinese | WPRIM | ID: wpr-808649

ABSTRACT

Objective@#To analyze the etiological of herpangina(HA) in Guangzhou City in 2015, and to provide laboratory data for the epidemic control.@*Methods@#Two hundred and eleven herpangina samples (stool and throat swab) were collected.Real-time (RT)-PCR and semi-nested (Sn)-PCR assays were performed to detect human enteroviruses (HEVs)-positive samples. The human rhabdomyosarcoma (RDa) cell lines were used to inoculate virus from HEVs-positive samples. The entire sequences of viral genes encoding VP1 of CVA6 positive samples or strains were amplified and sequenced. The phylogenetic analysis was performed to analyze the full-length gene sequences encoding VP1 of CVA6 by using DNAStar6.0 and MEGA5.2 software packages.@*Results@#According to the laboratory test results, 115 cases were HEVs-positive and positive rate was 93.50%, eight serotypes of EV including CVA6, CVA10, CVA2, EV71, CVA16, CVB2, Echo14 and Echo30 were detected.The CVA6 positive rate was the highest with a percentage of 60.98%, followed by CVA10 with a percentage of 13.01%. The enterovirus positive rate of stool samples (χ2=29.88, P<0.01) and viruses isolated positive rate (χ2=8.67, P<0.01) were higher than that in throat swab samples. Phylogenetic analysis showed that all CVA6 strains detected in this study belonged to D3 subgenotype, and shared 96.9%-99.9% homologies in nucleotide and 99.0%-100.0% in amino acid.@*Conclusions@#CVA6 of the enterovirus A group accounted for the main pathogen of herpangina in Guangzhou City in Guangdong province in 2015, which belonged to D3 subgenotype.

8.
Chinese Journal of Experimental and Clinical Virology ; (6): 367-371, 2017.
Article in Chinese | WPRIM | ID: wpr-808488

ABSTRACT

Objective@#To develop a micro-neutralization test for determination of neutralizing antibody against ZIKA virus (ZIKV) in human sera and to verify the acute and convalescent serum samples of 10 ZIKA virus-infected cases diagnosed by nucleic acid detection and/or virus isolation.@*Methods@#ZIKV isolated from ZIKA cases was used to determine micro-neutralization antibody. The virus solution was prepared by infecting BHK21, VERO and VERO-E6 cell lines and viral titer was tested; 100 TCID50 viral solution and 4 times diluted sera which were inactivated at 56 ℃ for 30 min were neutralized, then added the cell suspension and incubated in 5% CO2 incubator at 37 ℃ for 7 d. The CPE was observed every day.@*Results@#The sensitivity of BHK21, VERO and VERO-E6 was different after infection with ZIKA virus. VERO cell line was the most sensitive and showed typical CPE. VERO cell line was used to establish a micro-neutralization test for determination of neutralizing antibody against ZIKA virus in sera.@*Conclusions@#The neutralizing antibody test for zika virus in sera is a special and usefulmethod to diagnose human infection of ZIKV and to conduct population based epidemiological investigation.

9.
Chinese Journal of Epidemiology ; (12): 1541-1545, 2017.
Article in Chinese | WPRIM | ID: wpr-737870

ABSTRACT

Objective To investigated the etiologic characteristics of Shigella (S.) sonnei strains causing outbreaks and sporadic cases in some areas of Guangdong province and Guangxi Zhuang Autonomous Region during 2014-2016. Methods Fourteen S. sonnei strains isolated from outbreaks and 6 S. sonnei strains from sporadic cases from Guangdong and Liuzhou of Guangxi Zhuang Autonomous Region were tested for antimicrobial resistance and analyzed by pulsed-field gel electrophoresis (PFGE). Six typical strains were selected for whole genome sequencing typing and compared with 51 strains isolated both at home and abroad from NCBI genome database. Results The antibiotic resistance test indicated the isolates had high resistance rate to ampicillin, tetracycline, gentamicin, trimethoprim/sulfamethoxazole and nalidixic acid, while sensitive to azithromycin, chloromycetin and imipenem. PFGE showed high similarity (93.2%) among the strains isolated from different areas. The whole genome sequencing analysis also revealed that all the typical strains wereclustered into a same evolution branch, close to some strains from Korea. Conclusions The S. sonnei strains isolated from some areas of Guangdong and Guangxi Zhuang Autonomous Region showed high resistance to commonly used antibiotics, but they were sensitive to azithromycin, chloramphenicol and imipenem. The isolates in this study also showed similar PFGE patterns and close phylogenic evolution.

10.
Chinese Journal of Epidemiology ; (12): 906-910, 2017.
Article in Chinese | WPRIM | ID: wpr-737745

ABSTRACT

Objective To analyze transmission factors of norovirus outbreaks in Guangdong province during 2008-2015 and provide evidence for the prevention and control of norovirus infection.Methods Epidemiological analysis was performed on the data of norovirus outbreaks reported in Guangdong from January 1,2008 to December 31,2015,which were obtained from the Public Health Emergency Management Information System of Guangdong province.The samples collected from the norovirus outbreaks were detected for norovirus by RT-PCR and the gene sequencing of the positive PCR products were performed.Results A total of 96 norovirus outbreaks were reported in Guangdong during 2008-2015.Sixteen outbreaks were reported during 2008-2012and 80 outbreaks were reported during 2013-2015 (83.3%).Eighty-two outbreaks (85.4%) occurred in schools.The infection routes included foodborne transmission in 39 outbreaks (40.6%),person to person transmission in 23 outbreaks (24.0%) and waterborne transmission in 8 outbreaks (7.3%).The gene sequencing results showed that variant G Ⅱ.4/Sydney2012 was the predominant pathogen for 6 of the 20 outbreaks (30.0%) during 2012-2013.Variant G lⅡ.17 was the predominant pathogens for 33 of the 53 outbreaks (62.3%) during 2014-2015.Conclusion The norovirus outbreaks in Guangdong during 2008-2015 were caused by foodborne and person to person transmissions of two emerging variant:G Ⅱ.4/Sydney2012 and G Ⅱ.17.

11.
Chinese Journal of Epidemiology ; (12): 1541-1545, 2017.
Article in Chinese | WPRIM | ID: wpr-736402

ABSTRACT

Objective To investigated the etiologic characteristics of Shigella (S.) sonnei strains causing outbreaks and sporadic cases in some areas of Guangdong province and Guangxi Zhuang Autonomous Region during 2014-2016. Methods Fourteen S. sonnei strains isolated from outbreaks and 6 S. sonnei strains from sporadic cases from Guangdong and Liuzhou of Guangxi Zhuang Autonomous Region were tested for antimicrobial resistance and analyzed by pulsed-field gel electrophoresis (PFGE). Six typical strains were selected for whole genome sequencing typing and compared with 51 strains isolated both at home and abroad from NCBI genome database. Results The antibiotic resistance test indicated the isolates had high resistance rate to ampicillin, tetracycline, gentamicin, trimethoprim/sulfamethoxazole and nalidixic acid, while sensitive to azithromycin, chloromycetin and imipenem. PFGE showed high similarity (93.2%) among the strains isolated from different areas. The whole genome sequencing analysis also revealed that all the typical strains wereclustered into a same evolution branch, close to some strains from Korea. Conclusions The S. sonnei strains isolated from some areas of Guangdong and Guangxi Zhuang Autonomous Region showed high resistance to commonly used antibiotics, but they were sensitive to azithromycin, chloramphenicol and imipenem. The isolates in this study also showed similar PFGE patterns and close phylogenic evolution.

12.
Chinese Journal of Epidemiology ; (12): 906-910, 2017.
Article in Chinese | WPRIM | ID: wpr-736277

ABSTRACT

Objective To analyze transmission factors of norovirus outbreaks in Guangdong province during 2008-2015 and provide evidence for the prevention and control of norovirus infection.Methods Epidemiological analysis was performed on the data of norovirus outbreaks reported in Guangdong from January 1,2008 to December 31,2015,which were obtained from the Public Health Emergency Management Information System of Guangdong province.The samples collected from the norovirus outbreaks were detected for norovirus by RT-PCR and the gene sequencing of the positive PCR products were performed.Results A total of 96 norovirus outbreaks were reported in Guangdong during 2008-2015.Sixteen outbreaks were reported during 2008-2012and 80 outbreaks were reported during 2013-2015 (83.3%).Eighty-two outbreaks (85.4%) occurred in schools.The infection routes included foodborne transmission in 39 outbreaks (40.6%),person to person transmission in 23 outbreaks (24.0%) and waterborne transmission in 8 outbreaks (7.3%).The gene sequencing results showed that variant G Ⅱ.4/Sydney2012 was the predominant pathogen for 6 of the 20 outbreaks (30.0%) during 2012-2013.Variant G lⅡ.17 was the predominant pathogens for 33 of the 53 outbreaks (62.3%) during 2014-2015.Conclusion The norovirus outbreaks in Guangdong during 2008-2015 were caused by foodborne and person to person transmissions of two emerging variant:G Ⅱ.4/Sydney2012 and G Ⅱ.17.

13.
Chinese Journal of Epidemiology ; (12): 1137-1141, 2016.
Article in Chinese | WPRIM | ID: wpr-737543

ABSTRACT

Objective To understand the effect ofserotyping on Salmonella isolates,by use of Microsphere-based Liquid Array method,among diarrhea patients,in Guangdong.Methods Salmonella isolated from humans in Guangdong province were serotyped on the Microsphere-based Liquid Array platform with SSA kit.Results A total of 4 942 Salmonella strains with 189 serotypes,were identified in Guangdong province in 2010-2014.The top 100 serotypes accounted for 98.08% (4 847/4 942) of all the strains.98% of the top 100 species serotypes could completely be serotyped with SSA kit.In order to detect O antigen among 198 isolates with SSA kit,181 strains were carrying the O antigen,with the coincidence rate as 100%.However,under the SSA,98.32% (528/537) of the H antigen could be detected and were consistent with the traditional serum agglutination test.The coincidence rate of fljB gene was 93.09% (175/188),with false negative rate and false positive rate of fljB gene as 7.35% (9/134) and 7.41% (4/54) respectively.The coincidence rate of sdf gene and Vi gene were 100%.11 out of the 12 Salmonella strains could not be serotyped under the traditional methods but were successfully serotyped by the molecular serotyping method.Conclusions Using the SSA kit,more than 96% of the anthropogenic Salmonella strains could be serotyped in Guangdong province.Comparing with the traditional methods,the coincidence rate of serotyping appeared over 98%.Under the Microsphere-based Liquid Array techniques,the molecular serotyping method appeared faster and more accurate on Salmonella serotyping than those traditional methods.

14.
Chinese Journal of Epidemiology ; (12): 1137-1141, 2016.
Article in Chinese | WPRIM | ID: wpr-736075

ABSTRACT

Objective To understand the effect ofserotyping on Salmonella isolates,by use of Microsphere-based Liquid Array method,among diarrhea patients,in Guangdong.Methods Salmonella isolated from humans in Guangdong province were serotyped on the Microsphere-based Liquid Array platform with SSA kit.Results A total of 4 942 Salmonella strains with 189 serotypes,were identified in Guangdong province in 2010-2014.The top 100 serotypes accounted for 98.08% (4 847/4 942) of all the strains.98% of the top 100 species serotypes could completely be serotyped with SSA kit.In order to detect O antigen among 198 isolates with SSA kit,181 strains were carrying the O antigen,with the coincidence rate as 100%.However,under the SSA,98.32% (528/537) of the H antigen could be detected and were consistent with the traditional serum agglutination test.The coincidence rate of fljB gene was 93.09% (175/188),with false negative rate and false positive rate of fljB gene as 7.35% (9/134) and 7.41% (4/54) respectively.The coincidence rate of sdf gene and Vi gene were 100%.11 out of the 12 Salmonella strains could not be serotyped under the traditional methods but were successfully serotyped by the molecular serotyping method.Conclusions Using the SSA kit,more than 96% of the anthropogenic Salmonella strains could be serotyped in Guangdong province.Comparing with the traditional methods,the coincidence rate of serotyping appeared over 98%.Under the Microsphere-based Liquid Array techniques,the molecular serotyping method appeared faster and more accurate on Salmonella serotyping than those traditional methods.

15.
Chinese Journal of Microbiology and Immunology ; (12): 506-511, 2016.
Article in Chinese | WPRIM | ID: wpr-495756

ABSTRACT

Objective To prepare simulated stool specimens for proficiency testing ( PT) by mix-ing lentils with Salmonella, Shigella and Escherichia coli strains and to establish an assessment scheme for the detection of Salmonella and Shigella in clinical samples. Methods Salmonella, Shigella and Escherich-ia coli strains were respectively spiked to lentils in Cary-Blair transport medium to create simulated stool specimens. Various ratios of Escherichia coli to Salmonella strains were spiked to lentils to prepare mixed simulated stool specimens. The accuracy and stability of prepared stool samples for PT were tested in-house. Results of sample detection were collected from participating laboratories for further external quality assess-ment. Results The Escherichia coli and Salmonella strains mixed at ratios of 100 ∶ 1 to 106 ∶ 1 could be ef-ficiently isolated from the media. Enrichment was needed in order to effectively isolate Salmonella strains from the media when the ratios of Escherichia coli to Salmonella strains were 104 ∶ 1 to 106 ∶ 1. Of the16 participating laboratories, 14 laboratories (87. 5%) received a grade of“satisfactory” and the other 2 labo-ratories (12. 5%) received a grade of “mainly satisfactory”. Conclusion The simulated stool specimens and the PT procedures designed in this study were suitable for proficiency testing program on the detection of Salmonella, Shigella and other similar microbes.

16.
Chinese Journal of Microbiology and Immunology ; (12): 247-251, 2016.
Article in Chinese | WPRIM | ID: wpr-486739

ABSTRACT

Objective To establish a method for the isolation of Zika virus and to gather experi-ences for viral isolation. Methods Suckling mice at age 1-3 days were inoculated with serum samples posi-tive for Zika virus through intracranial injection. All mice were sacrificed 6 days after the injection. Viral nu-cleic acids were extracted from brain, heart, liver, spleen, lung, kidney, muscle, skin and intestine tissue samples and analyzed by real-time RT-PCR. The supernatants of brain tissues positive for Zika virus were used for subculturing. Nested PCR was performed to amplify the NS5 gene of the isolated virus. The se-quences of NS5 gene were analyzed by using MEGA6. 0 software. Results All of the tissue samples were positive for Zika virus. Higher viral loads were detected in heart and brain tissue samples with cycle thresh-old (Ct) values of 24. 4 and 25. 3, respectively. The second generation of Zika virus was identified in suck-ling mice brain tissues 2 days after infection by using real-time RT-PCR. The amplified product of nested PCR was 972 bp in length. Sequencing analysis showed that the isolated Zika virus ( GDZ16002 strain) be-longed to the Asian lineage. Conclusion A strain of Zika virus was successfully isolated in China by using intracranial injection via a suckling mouse model. The isolated Zika virus belonged to the Asian lineage.

17.
Chinese Journal of Microbiology and Immunology ; (12): 93-97, 2016.
Article in Chinese | WPRIM | ID: wpr-483755

ABSTRACT

Objective To establish an inactivated viral particle-based ELISA for the detection of antibodies against Middle East respiratory syndrome coronavirus (MERS-CoV) in serum samples collected from a MERS-CoV associated case. Methods Serum samples were collected from 10 newborns and 40 healthy adults. A viral particle-based ELISA was established by using the inactivated MERS-CoV virions as antigen. The levels of IgM and IgG antibodies in the serum samples were detected by the established ELISA and the cut-off values for positive detection were determined. Then the inactivated MERS-CoV virion-based ELISA was used to detect the antibodies against MERS-CoV in 5 serum samples collected from the first im-ported MERS case in China. Results The cut-off values of IgM and IgG antibodies in serum samples for ELISA were determined to be A450 readings of 0. 32 and 0. 42, respectively. The titers of IgM and IgG anti-bodies in serum samples collected at early admission to hospital from the first imported MERS case in China were both 1 ︰ 40. Seroconversion occurred 2 weeks after his admission to hospital with the titers of IgM and IgG reaching to 1 ︰ 320. Conclusion The inactivated MERS-CoV virion-based ELISA was established successfully and could be used for the detection of serum antibodies (IgG and IgM) in MERS associated cases.

18.
Chinese Journal of Preventive Medicine ; (12): 615-620, 2015.
Article in Chinese | WPRIM | ID: wpr-270027

ABSTRACT

<p><b>OBJECTIVE</b>To analyze epidemiological characteristics of norovirus variant of GII.4 Sydney from January 2012 to June 2014 in sentinel hospitals of Guangdong province, as well as the outbreaks caused by norovirus variant of GII.4 Sydney.</p><p><b>METHODS</b>During January 2012 to June 2014, a total of 10 750 fecal samples were obtained from 22 hospitals of surveillance sites in Guangdong province. Those samples were sent to the local municipal CDCs for extracting and detecting norovirus nucleic acid. Then, all the positive samples were delivered to Guangdong provincial CDC that used Random Number Method to draw 855 positive samples for norovirus genotyping, and 690 samples were successfully sequenced. Chi-square tests were used to compare norovirus infection status of diarrhea cases in different age groups as well as during different periods. Epidemiological data of 13 outbreaks which were caused by norovirus variant of GII.4 Sydney from January 2012 to June 2014 were collected from the Public Health Emergency Management Information System of Guangdong Province, and the epidemiological characteristics were analyzed.</p><p><b>RESULTS</b>The norovirus variant of GII.4 Sydney was first detected in August 2012 and the detection rate was 13/15 in November 2012. During November 2012 to January 2013 (period T1), the norovirus positive rate of each month was 23.8% (100/421), 15.9% (61/383) and 19.2% (95/495), respectively. During November 2013 to January 2014 (period T2), the norovirus positive rate of each month was 17.0% (90/529), 8.7% (37/426) and 11.2% (46/409), respectively which were significantly lower than that of period T1 (χ² alue was 6.65, 9.93 and 10.74. P value was 0.010, 0.002, and 0.001, respectively). In period T1, the norovirus positive rate of people ages 15 and older was 26.3% (143/543) and the rate of people under 15 was 14.9% (113/756) (χ² = 2.90, P < 0.001). In period T2, the norovirus positive rate of people ages 15 and older was 10.1% (52/516) and the rate of people under 15 (14.3% (121/848)) (χ²= 5.09, P = 0.024). The foodborne transmission was the infection source for ten of thirteen outbreaks.</p><p><b>CONCLUSION</b>The norovirus variant of GII.4 Sydney was first detected in August 2012. The epidemic began to occur in the community since November 2012, and the strength of the epidemic declined 1 year later. The foodborne transmission was the main infection sources for the outbreaks caused by norovirus variant of GII.4 Sydney.</p>


Subject(s)
Adolescent , Adult , Child , Humans , Chenodeoxycholic Acid , China , Diarrhea , Disease Outbreaks , Epidemics , Foodborne Diseases , Genotype , Norovirus , Sentinel Surveillance
19.
The Journal of Practical Medicine ; (24): 999-1002, 2015.
Article in Chinese | WPRIM | ID: wpr-464694

ABSTRACT

Objective To study the factors influencing epidemiological characteristics and virulence of Enterovirus 71 (EV71) in Guangdong province from 2008 to 2010. Methods RNA was extracted from collected samples or cultured virus , then reversing transcription into cDNA. We amplified full-length EV71-VP1 using poly-merase chain reaction technology , then conducted sequence alignment and established phylogenetic tree with MEGA software (version 5.0) to confirm the genotype of EV71. The association between severity of clinical symp-toms and sex, age, viral genotype and VP1 variation was also analyzed using Logistic regression. Results The genotype of the predominant epidemical strain was C4a in Guangdong from 2008 to 2010. However , this subtype had already differentiated into 4 subgroups (C4a1- C4a4). There was no correlation between clinical syndrome and sex or viral genotype; the severity of symptoms was negatively correlated with age: before 4 years old, varia-tion A289T can easily lead to severe cases, increasing the risk of infection (P<0.05, OR = 2.360, 95%CI:1.163~ 4.659). Conclusion The main epidemical EV71 strain is C4a1 in Guangdong province. The emerging differen-tiation and simultaneous prevalence should merit attention to strengthen relevant surveillance; and the protection of the susceptible population should be reinforced during EV71 prevalence.

20.
Chinese Journal of Virology ; (6): 333-340, 2015.
Article in Chinese | WPRIM | ID: wpr-296279

ABSTRACT

To investigate the genetic character and origin of the first imported infection case of middle East respiratory syndrome coronavirus (named as MERS-CoV_China GD01), RNA was extracted from swabs of this patient followed by RT-PCR amplification. All coding gene of structural (S, E, M, E) and accessory (ORF3, ORF4a, ORF4b, ORF5, ORF8b) proteins were sequenced and analyzed. Phylogenetic analyses of structural protein coding genes of MERS-CoV_ China GD01 indicates that several substitutes exists in S coding gene and its origin belong group 5 of MERS-CoV, which were recent circulated in Saudi Arabia area, while other three structural genes (N, E, M) were very conserved. Phylogenetic analyses of accessory protein coding genes of MERS-CoV China GD01 indicates that several substitutes exists among ORF3, ORF4a, ORF4b and ORF5, while ORF8b was conserved. In conclusion, genome of MERS-CoV_ China GD01 was general conserved although several genetic variations were found among structural and accessory protein coding genes. This is the first report on sequencing and phylogenetic analyses of the first imported MERS case in China, which may pay the way for prevention and control of imported MERS-CoV infection.


Subject(s)
Humans , China , Conserved Sequence , Coronavirus Infections , Virology , Evolution, Molecular , Genomics , Middle East Respiratory Syndrome Coronavirus , Genetics , Physiology , Phylogeny , Sequence Analysis , Viral Proteins , Genetics
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