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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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Objective To investigate the serotypes, antimicrobial resistance, virulence genes and molecular characteristics of food-borne Vibrio parahaemolytieus isolated in Guangdong province in 2014.Methods Sixty V.parahaemolyticus strains were tested in this study.The serotyping and antibiotic resistance test were conducted, while the tdh and trh gens of the strains were detected with conventional PCR, and pulsed field gel electrophoresis (PFGE) and multiple locus sequence typing (MLST) were conducted too.Results The 60 strains belonged to 13 serotypes, the major serotypes included O3 : K6, O4: K8, O 1 : K36 and O4: KUT.The antibiotic resistance test indicated the isolates were highly resistant to ampicillin (100.0％), sulfonamides (43.3％) and cefalotin (28.3％).Up to 56.7％(34/60) of the strains were resistant to two or more antibiotics, and 2 strains showed resistance to three antibiotics.The virulence gene detection indicated that 63.3％ (38/60) of the strains carded tdh+trh-, while only 1 strain carried tdh+trh+.The 60 strains digested by Not I belonged to 48 different PFGE patterns and 3 clusters.The cluster B included the strains isolated from sporadic food borne cases with serotype of O3:K6 and similarity of 62.6％-100.0％.The cluster C included O4:K8 strains with the PFGE pattem similarity of 56.7％-62.5％.The MLST indicated that the 60 strains had 26 sequence types (STs).The ST-3 was predominant, including 33 strains (serotypes O3:K6 and O1 : K36).The four O4:K8 strains formed another predominant colony, which was different from ST-3.Conclusion The etiologic characteristics of V.parahaemolyticus varied, which might be one of the reasons for high incidence of food-bome V.parahaemolyticus infection in Guangdong.The molecular characteristics of O4: K8 strain were differem from the other predominant serotypes.Close attention should be paid to the possible outbreak caused by O4: K8 strain in this area.
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<p><b>OBJECTIVE</b>To analyze the etiologic characteristics of O1/O139 Vibrio cholerae in Guangdong province in 2009-2013.</p><p><b>METHODS</b>Isolates from cholera cases and from the environment surveillance points were investigated by serological typing, antibiotic susceptibility testings, toxic genes detection and molecular typing to analyze the similarities and differences of the identified species.</p><p><b>RESULTS</b>Totally, 190 isolations of O1/O139 V. cholerae were obtained from cholera cases (16 strains) and environmental samples (174 strains) in Guangdong province in 2009-2013. The sero-types would include Inaba (3 isolates), Ogawa (7 isolates) and O139 (6 isolates) in all the isolates from the cholera cases. Ten strains from the ctxA positive cases were detected by PCR. Two Ogawa strains carried incomplete CTXΦ phage. Results from the antibiotic susceptibility test indicated that 5 strains were absolutely sensitive to 11 antibiotic discs in vitro, while another 3 strains were simultaneously resistant to 4 antibiotic discs. Except for 2 stains, all the O139 strains from the environment were ctxA negative, detected by PCR. Incomplete CTXΦ phage was found in the Inaba (53 isolates), Ogawa (22 isolates) and O139 (2 isolates), respectively. Results from the antibiotic susceptibility test exhibited that 25 strains were resistant simultaneously to 4 and/or more antibiotic discs, especially the Inaba strains from the seafoods(13 isolates). 2 Inaba strains from seafood were simultaneously resistant to 7 antibiotic discs. Results from PFGE molecular typing indicated that the PFGE types digested by Not I expressed significant diversity. Inaba and O139 strains from cases were gathered in the same clusters, while the Ogawa strains from cases scattered in different clusters but no significant correlation among these strains were found. Our results suggested that a distant genetic relationship might exist between these two different sources strains.</p><p><b>CONCLUSION</b>Complex and diverse as the virulence genes and genetic characteristics and with the grim situation of multi-drug resistant strains, all seemed important to strengthen the surveillance programs on the variation of strain types and antibiotics resistance of O1/O139 V. cholerae in Guangdong province.</p>
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Humanos , Técnicas de Tipagem Bacteriana , China , Epidemiologia , Cólera , Epidemiologia , Microbiologia , Farmacorresistência Bacteriana Múltipla , Genes Bacterianos , Testes de Sensibilidade Microbiana , Vibrio cholerae , Classificação , VirulênciaRESUMO
Objective To understand the distribution and the characteristics on molecular typing of Salmonella (S.) typhimurium isolates gathered from the surveillance program and to construct the standard S.typhimurium databank in the laboratory through surveillance network PulseNet,in Guangdong province to improve the capability of detection on laboratory-based foodbome outbreaks.Methods With the application of standard pulse-field gel electrophoresis (PFGE) and multiple loci VNTR analysis (MLVA) including seven VNTRs loci protocols on PulseNet International Network,275 isolates of S.typhimurium from ten cities in Guangdong province were typed and their patterns analyzed.The molecular typing databank was constructed by BioNumerics.Results With S.typhimurium the most common serotypes,the average annual positive rate of Salmonella strains and S.typhimurium were 4.03％ and 1.38％ respectively.The positive rate and proportion presented a double-peak trend,appearing in May and September.The chromosomal DNA of S.typhimurium was digested with Xba Ⅰ restricted endonucleotidase and 124 PFGE patterns were observed after pulse-field gel electrophoresis,with the discrimination index (D) as 0.928 6.The patterns including more than three S.typhimurium isolates and were further digested with the second enzyme Bln Ⅰ to achieve 174 patterns,with the D value as 0.989 1.Under MLVA method,143 variant patterns were obtained,with the D value reaching 0.966 5.Data showed that the discriminatory ability of the MLVA typing method in S.typhimurium was superior to PFGE-Xba Ⅰ but equal to PFGE-Xba Ⅰ-Bln Ⅰ.In addition,when S.typhimurium strains were respectively analyzed by PFGE under double enzymes digestion and MLVA,the results appeared coincident and relative.Conclusion The variant patterns showed by the two molecular typing methods indicating a genetic diversity existed among the clinical S.typhimurium isolates in Guangdong province.Databank of S.typhimurium was constructed and could be used in laboratory surveillance programs.Under the characterization of analyzing similarity and evolution among S.typhimurium isolates,MLVA was suitable for cluster analysis on early detection of outbreaks caused by S.typhimurium.
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<p><b>OBJECTIVE</b>To study the serotypes, antimicrobial resistance, virulence genes and molecular characteristics of Vibrio parahaemolyticus isolated from outbreaks and sporadic cases in Guangdong, 2013.</p><p><b>METHODS</b>36 Vibrio parahaemolyticus strains isolated from outbreaks and 43 strains from sporadic cases were sero-typed and tested for antimicrobial resistance. PCR was used to detect for tdh(thermostable direct hemolysin gene), trh (tdh(-) related hemolysin gene), GS-PCR and orf8 genes. All the samples were analyzed by pulsed-field gel electrophoresis (PFGE).</p><p><b>RESULTS</b>36 isolates from outbreaks were all identified as O3 : K6, and among the 43 sporadic isolates, O3 : K6 (23, 53.49%) was the dominant serotype. Vibrio parahaemolyticus isolates showed high resistance rate to ampicillin (96.20%) and cefalotin (40.50%), but were high sensitive to cotrimoxazole (100%) and chloramphenicol (100%). 83.33% (30/36) outbreak isolates were resistant to multi-drugs but only 37.21% (16/43) of the sporadic isolates showed so. Results from virulence gene detection suggested that all the 36 outbreak isolates belonged to tdh(+) trh(-) strains, while 86.05% (37/43) of the sporadic isolates were tdh(+)trh(-) and 11.63% (5/43)were tdh(-)trh(+) . Only one tdh(+)trh(+) strain was found. All the outbreak isolates contained GS-PCR and/or orf8 genes, whereas among the sporadic isolates only 51.16% (22/43) of them carrying the similar genes. Results from PFGE analysis suggested that 79 isolates were discriminated into 3 clusters and 32 different PFGE patterns with the similarity value between 59.8% and 100.0%. Outbreak isolates seemed to gather in the same cluster, while the sporadic isolates spreading in all the three clusters.</p><p><b>CONCLUSION</b>O3 : K6 was the dominant serotype in Vibrio parahaemolyticus strains isolated in Guangdong, 2013. These strains showed high sensitivity to most antibiotics, but with multi-drug resistance. Positive rate of tdh gene was high, and most O3 : K6 strains contained GS-PCR and/or orf8 genes. PFGE analysis revealed genetic diversity was within the Vibrio parahaemolyticus strains in Guangdong.</p>
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Humanos , Toxinas Bacterianas , China , Epidemiologia , Surtos de Doenças , Farmacorresistência Bacteriana , Eletroforese em Gel de Campo Pulsado , Variação Genética , Proteínas Hemolisinas , Reação em Cadeia da Polimerase , Sorotipagem , Vibrioses , Epidemiologia , Vibrio parahaemolyticus , Genética , Virulência , VirulênciaRESUMO
To compare and evaluate the discriminatory ability and potential value of pulsed field gel electrophoresis (PF-GE) and multiple locus VNTRs analysis (MLVA) on the genotyping of Brucella ,a total of 60 strains of Brucella and three standards (16M ,544A ,1330S) were genotyped simultaneously by PFGE and MLVA .The result indicated that the similarity coefficient among the 63 isolates was from 72 .1-100 .0% by PFGE ,and could distinguish three species of B .melitensis ,B .su-is and B .abortus at the similarity level of 94 .4% .There were 14 clusters and 29 PFGE types identified by PFGE with discrim-inatory index (DI) of 0 .957 5 at the similarity level of 100% ;the similarity coefficient among the 63 isolates was from 16 .9-100 .0% by MLVA ,and could distinguish three species of Brucella at the similarity level of 52 .3% .There were 8 clusters and 47 MLVA types identified by MLVA with discriminatory index (DI) of 0 .985 2 at the similarity level of 100% .It's suggested that PFGE and MLVA could be used to distinguish three species of Brucella in the similarity coefficient of certain ,but could not effectively distinguish the type in the same species .Both of these two methods could be used for Brucella molecular typing , but MLVA is better than PFGE for its relatively higher discriminating ability .
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Objective To investigate the etiological characteristics of Streptococcus pyogenes that caused scarlet fever in different periods in Guangdong province.Methods 22 isolates from different periods were analyzed through emm typing,PCR detection for super antigen genes,antibiotic susceptibility test and pulsed-field gel electrophoresis (PFGE).Results All isolates were susceptible to cefotaxim,levofloxacin and penicillin.Streptococcus pyogenes isolated after the year 2000 were 100％ resistant to erythrocin and clindamycin,but the resistant rate for strains isolated before the year 2000 was 9.1％ (1/11).There were 3 emm types indentified from 22 isolates including emm12.0 (59.09％,13/22),emm6.0 (36.36％,8/22) and emm1.0 (4.55％,1/22),which were detected in the isolates from the year 1997 and 2011,from 1978 and 1986,and from 2008,respectively.The positive rates for speA,speB,speC,speF,speG,speH,smeZ,and ssa genes detected by PCR were 54.55％,100％,100％,100％,100％,54.55％,0％,and 86.36％ respectively.Among all strains,95.45％ of the isolates carried 6 superantigen genes simultaneously.Three clusters of 10 PFGE subtypes were identified in 22 isolates.Cluster Ⅰ consisted of all strains from 1997 and one strain from 2011.Cluster Ⅱ consisted of strains isolated from 1978 and 1986.Cluster Ⅲ consisted of nine strains from 2011 and one from 2008.Conclusion S.pyrogenes isolates in Guangdong province were susceptible to penicillin but resistant to erythrocin.emm 12.0 accounted for the majority of the three types and there was a high frequency of super antigen genes.
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Objective To investigate the polymorphism of Brucella melitensis biovar 3 ( B.melitensis biovar 3) strains isolated from Guangdong province .Methods PCR assays followed by agar-ose gel electrophoresis and capillary electrophoresis based on the multiple locus variable number of tandem repeats (VNTR) analysis (MLVA) were performed to analyze 43 clinic isolates of B.melitensis biovar 3 strains isolated from clinical patients in Guangdong province .Results MLVA typing showed that the simi-larities of the analyzed locus among 43 strains of Brucella ranged from 68.2%to 100%.32 genotypes identi-fied among the isolates were identical (100%similarity).27 out of 43 strains (62.8%) were single geno-types, while the other 16 strains (37.2%) belonged to 5 other genotypes with 2 to 5 strains in each of them . Conclusion B.melitensis biovar 3 isolates showed polymorphism distribution in Guangdong province as in-dicated by MLVA typing analysis .Single-genotype isolates accounted for 62.8% of all studied strains.No predominant genotype was found among all isolates .
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Objective To investigate the antimicrobial resistance pattern of non-typhoidal Salmonella isolated from diarrhea cases in Guangdong province,China.The multidrug-resistant strains were analyzed by pulsed field gel electrophoresis(PFGE) typing.Methods All the non-typhoidal Salmonella strains isolated between 2009 and 2011 were serotyped,then the antimicrobial resistance was detected by the disk diffusion method and molecular typed by PFGE.Results 91.76％ (256/279) S.typhimurium isolates were multiple resistant to 3 and more antimicrobials.Forty S.typhimurium isolates were multiple resistant to 9 and more antimicrobials and 3 out of which were multiple resistant to all the 12 antimicrobials in vitro.96.91％ (94/97) Salmonella I4,5,12:i:-isolates were multiple resistant to 3 and more antimicrobials.Nine Salmonella I4,5,12:i:- isolates were multiple resistant to 9 and more antimicrobials and I out of which was multiple resistant to all the 12 antimicrobials1 in vitro.47％ (47/100) S.enteritidis isolates were multiple resistant to 3 and more antimicrobials.Only 1 S.enteritidis isolates was multiple resistant to 9 and more antimicrobials.4.27％ (27/632) non-typhoidal Salmonella isolates was resistant to ciprofloxacin,including 17 S.typhimurium and 6 Salmonella 14,5,12:i:- isolates.Also,there were 3 1.96％ ( 202/632 ) non-typhoidal Salmonella isolates was intermediary to ciprofloxacin.The PFGE patterns of the predominant strains which were highly resistant and multidrug-resistant had different genotypes and demonstrated significant genetic diversity.Conclusion The situation about the multiple antimicrobial resistances of non-typhoidal Salmonella in Guangdong province has showed the prevalent problem.The PFGE types of the multiple drug-resistant strains prompted these strains were come from different clones.This requires that we continue to strengthen the resistance monitoring and control of the rational use of antibiotics.
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ObjectiveTo study the toxin genes and pandemic group distribution as well as the genetic correlation between the major serotypes( O3:K6,O1:Kut,O4:K8 ) of Vibrio parahaemolyticus (VP) isolated from the outbreaks of Guangdong province.MethodsThe tdh and trh genes,GS-PCR and orf8 gene were detected on the 62 isolates sourced from patient and seafood occurred in the 23 outbreaks during 2008-2010.44 isolates of which were analyzed on PFGE digested by Not Ⅰ enzyme.ResultsToxin genes distributions suggested that 96.8% (60/62)isolates were tdh+,trh-.Three tdh+ isolates sourced from seafood were found.Pandemic group distribution suggested that 97.2% (35/36) O3:K6,5.88% (1/17) O4:K8,66.7% (8/9)O1:Kut serotype was GS-PCR+ and/or orf8+,respectively.PFGE analysis suggested that 44 isolates were separated into 3 clusters,of which the similarity of PFGE profile was 80.5% in the pandemic group cluster constructed by 28 isolates,the similarity between pandemic group and non-pandemic group was 59.5%.Pandemic group of O3:K6,O1:Kut as well as 04:K8 isolated on some outbreaks were processing the same PFGE profiles.ConclusionThe characteristic of toxin genes of major serotypes VP isolated in the outbreaks of Guangdong province form 2008-2010 was tdh- present and trh- absent.Within the pandemic group,O3:K6 and O1:Kut were the major serotypes.In single outbreak,isolates belongs to pandemic group but with different serotype seem to be close correlations.
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Objective To develop a rapid and sensitive DNA microarray for Listeria monocytogenes detection.Methods A DNA microarray was developed using gyrB,ISR,16S rRNA,23S rRNA,hlyA,iap and prfA as the target genes and tested against 18 different species of known reference for repeatability,sensitivity,and specificity to verify the effectiveness of the chip.Results After testing of samples by the LM array,results show that the 70 mer Oligos synthesized by IDT are superior to the Oligos synthesized by Sagon with respect to both probe spotting or samples detection.The comparison of 3 spotting probe concentrations of 10 μmol/L,40 μmol/L and 80 μmol/L demonstrated that the 10 pmol/L probes result in good detection signals equivalent to the 40 μmol/L and 80 μmol/L probes.The repeatability and sensitivity evaluated by sample testing on the LM array revealed that the chips developed in this study have good repeatability and the lower limit of sample detection is 0.9 ng DNA.The LM array can distinguish clearly and definitively between Listeria and non-Listeria bacteria in the sample.Conclusion The microarray is able to rapidly detect and identify Listeria monocytogenes.
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To explore the possibility to type the Brucella strains isolated in Guangdong province with analytical method to detect the fatty acid components and to collect the basic data of fatty acid components of Brucella strains, 29 strains of Brucella were selected for analysis on the bacterial fatty acid components and the cluster analysis on the collected data was performed with Sherlock analysis soft-ware (MIDI). It was demonstrated that the main fatty acid components of Brucella strains isolated in Guangdong province were 19∶0 cycloω8c acid, 16∶0 acid and 18∶0 acid. The content of 19∶0cycloω8c acid was highest in B.abortus, followed by B.melitensis and lowest in B.suis.-In addition, the content differences of 19∶0cycloω8c and 18∶0 acid between B. melitensis and Brucella suis were statistically significant; and that of 19∶0cycloω8c and 18∶0 acid between strains isolated in 1965 and those isolated in recent 3 years was statistically significant. It was also shown that the fatty acid components of Brucella strains were stable, but the contents of fatty acid components were different in different species.-It is evident that at certain euclidean distance, 3 species of Brucella can be differentiated in species level.