1.Investigation on vibrio cholera carried in aquatic products of littoral areas, Zhejiang Province.
Hua-kun LÜ ; En-fu CHEN ; Shu-yun XIE ; Cheng-liang CHAI ; Yu-dong WEI ; Shun-tang MO ; Ju-lian YE ; Yun LUO
Chinese Journal of Preventive Medicine 2006;40(5):336-338
OBJECTIVETo examine vibrio cholera (V.C) in aquatic products of littoral area, Zhejiang Province and to provide scientific evidence for administration of aquatic products and cholera epidemic control.
METHODSAll 990 samples of aquatic products collected from local markets, eateries and aquafarms in three chosen areas. Samples were proliferated in alkaline liquid medium, and purified in NO: 4 medium, the isolations were identified biochemically, and phenotype of strains were defined by phagocyte and coagulation with V.C. diagnostic serum. Three virulence genes (ctx, ace, zct) of the isolated strains were detected by polymerase chain reaction (PCR).
RESULTSThere were 1.41% samples caught by V.C., having a carrying rate highest in turtles of 8.9%. 14 strains were defined as three serogroups, and the numbers of Inaba, Ogawa, and Hikojima types were 2, 2, 10 respectively. Virulence genes had detected in 9 of 12 stains. All genes were detected in 5 strains, only ZOT genes in 3 strains, and both CTX and ACE genes in 1 strain.
CONCLUSIONSAquatic products from inshore in Zhejiang Province caught with V.C. strains might be divided into three serogroups. Most of them should be virulence genes. Cholera epidemic outbreak might be caused by those contaminated products.
China ; Food Microbiology ; Genes, Bacterial ; Seafood ; microbiology ; Vibrio cholerae ; genetics ; isolation & purification ; Virulence Factors ; genetics
3.Study on the distribution of Yersinia enterocolitica in Nantong, Jinagsu Province.
Ling GU ; Hua WANG ; Feng-cai ZHU ; Zhi-yang SHI ; Xiao-yan ZENG ; Zhao-ying TAN ; Yu-chun XIAO ; Hai-yan QIU ; Xin WANG ; Dong JIN ; Zhi-gang CUI ; Bing WANG ; Biao KAN ; Xin-sheng WANG ; Huai-qi JING ; Jian-guo XU
Chinese Journal of Epidemiology 2005;26(10):786-789
OBJECTIVETo study the distribution of Yersinia enterocolitica and its virulence factors in Nantong, Jiangsu.
METHODSYersinia strains were isolated from livestock and poultry. Conventional PCR was used to detect the virulence factors of all strains and strain 0:8 was analyzed by pulsed-field gel electrophoresis(PFGE).
RESULTSThe combined isolation rate of Yersinia enterocolitica from livestock and poultry was 31.06% and the gene distribution characters were: 39.57% of them were ail-, ystA- , ystB-, yadA- , virF-; 60.43% were ail- , ystA- , ystB + , yadA- , virF- respectively. The two reference strains from America and Denmark showed similar electrophoresis patterns but were significantly different with O:8 strains isolated from China while the serotypes of Yersinia enterocolitica O:3 and O:9 which were the main epidemic strains in China, were not found in this area.
CONCLUSIONThe pathogenic Yersinia enterocolitis O:3 and O:9 were not found in Nantong,Jiangsu province.
Animals ; Animals, Domestic ; microbiology ; China ; Electrophoresis ; Poultry ; microbiology ; Virulence Factors ; genetics ; metabolism ; Yersinia enterocolitica ; genetics ; isolation & purification ; pathogenicity
4.Detection of Yersinia Enterocolitica Bacteriophage PhiYe-F10 Lysis Spectrum and Analysis of the Relationship between Lysis Ability and Virulence Gene of Yersinia Enterocolitica.
Tao ZHA ; Junrong LIANG ; Yuchun XIAO ; Huaiqi JING
Chinese Journal of Virology 2016;32(2):185-189
To determine the lysis spectrum of Yersinia enterocolitica bacteriophage phiYe-F10 and to analyze the relationship between the lysis ability of phiYe-F10 and the virulence gene of Yersinia enterocolitica. To observe the lysis ability of the phage phiYe-F10 to the different Yersinia strains with the double-layer technique. The strains used in this study including 213 of Yersinia enterocolitica and 36 of Yersinia pseudotuberculosis and 1 of Yersinia pestis. The virulence genes of these Yersinia enterocolitica (attachment invasion locus (ail) and enterotoxin (ystA, ystB) and yersinia adhesin A (yadA), virulence factor (virF), specific gene for lipopolysaccharide O-side chain of serotype O : 3 (rfbc) were all detected. Among the 213 Yersinia enterocolitica, 84 strains were O : 3 serotype (78 strains with rfbc gene), 10 were serotype O : 5, 13 were serotype O : 8, 34 were serotype O : 9 and 72 were other serotypes. Of these, 77 were typical pathogenic Yersinia enterocolitica harboring with virulence plasmid (ail+, ystA+, ystB-, yadA+, virF+), and 15 were pathogenic bacterial strains deficiency virulence plasmid (ail+, ystA+, ystB-, yadA-, virF-) and the rest 121 were non pathogenic genotype strains. PhiYe-F10 lysed the 71 serotype O : 3 Yersinia enterocolitica strains which were all carried with rfbc+, including 52 pathogenic Yersinia enterocolitica, 19 nonpathogenic Y. enterocolitica. The phiYe-F10 can not lysed serotype O : 5, O : 9 and other serotype Y. enterocolitica, the lysis rate of serotype O : 3 was as high as 84.5%. The phiYe-F10 can not lysed Yersinia pseudotuberculosis and Yersinia pestis. Yersinia phage phiYe-F10 is highly specific for serotype O : 3 Yersinia enterocolitic at 25 degrees C, which showed a typical narrow lysis spectrum. Phage phiYe-F10 can lysed much more pathogenic Y. enterocolitica than nonpathogenic Y. enterocolitica.
Bacterial Proteins
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genetics
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metabolism
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Bacteriophages
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genetics
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isolation & purification
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physiology
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Host Specificity
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Virulence Factors
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genetics
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metabolism
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Yersinia enterocolitica
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genetics
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metabolism
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virology
5.Purification and characterization of a 33 kDa serine protease from Acanthamoeba lugdunensis KA/E2 isolated from a Korean keratitis patient.
Hyo Kyung KIM ; Young Ran HA ; Hak Sun YU ; Hyun Hee KONG ; Dong Il CHUNG
The Korean Journal of Parasitology 2003;41(4):189-196
In order to evaluate the possible roles of secretory proteases in the pathogenesis of amoebic keratitis, we purified and characterized a serine protease secreted by Acanthamoeba lugdunensis KA/E2, isolated from a Korean keratitis patient. The ammonium sulfate-precipitated culture supernatant of the isolate was purified by sequential chromatography on CM-Sepharose, Sephacryl S-200, and mono Q-anion exchange column. The purified 33 kDa protease had a pH optimum of 8.5 and a temperature optimum of 55 degrees C. Phenylmethylsulfonylfluoride and 4- (2- Aminoethyl) -benzenesulfonyl-fluoride, both serine protease specific inhibitors, inhibited almost completely the activity of the 33 kDa protease whereas other classes of inhibitors did not affect its activity. The 33 kDa enzyme degraded various extracellular matrix proteins and serum proteins. Our results strongly suggest that the 33 kDa serine protease secreted from this keratopathogenic Acanthamoeba play important roles in the pathogenesis of amoebic keratitis, such as in corneal tissue invasion, immune evasion and nutrient uptake.
Acanthamoeba/*enzymology/isolation & purification/pathogenicity
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Acanthamoeba Keratitis/*parasitology
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Animals
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Cornea/parasitology
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Humans
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Hydrogen-Ion Concentration
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Korea
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Serine Endopeptidases/chemistry/*isolation & purification/*metabolism
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Substrate Specificity
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Temperature
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Virulence Factors
6.Effect of solution environment on the purification of pertussis toxin.
Tie WU ; Jingxiu BI ; Yongdong HUANG ; Yan ZHANG ; Lijing SUN ; Chunbao SUN ; Zhiguo SU
Chinese Journal of Biotechnology 2008;24(7):1279-1284
The low recovery of pertussis toxin (PT) and the low resolving efficiency of chromatography, due to the instability of PT in low salt condition, are the main challenges for its purification. We aplied 2 mol/L urea to prevent the aggregation and disassociation of PT during the purification by ion-exchange chromatography (IEC) and gel filtration chromatography (GFC). The effect of urea on the purification of PT was studied by ELISA assay and non-reduced SDS-PAGE. The activity recoveries of PT and filamentous hemagglutinin (FHA) in IEC and GFC, the resolution efficiency in GFC and the purities of PT and FHA were improved obviously by adding 2 mol/L urea in the mobile phase. The results highlight the potential application of urea in the acellular pertussis vaccine (APV) manufacture procedure.
Adhesins, Bacterial
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isolation & purification
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Chromatography, Ion Exchange
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methods
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Humans
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Pertussis Toxin
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isolation & purification
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Pertussis Vaccine
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chemistry
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isolation & purification
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Solutions
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Urea
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chemistry
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Vaccines, Acellular
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chemistry
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isolation & purification
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Virulence Factors, Bordetella
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isolation & purification
7.Distribution of Virulence Genes in spa Types of Methicillin-resistant Staphylococcus aureus Isolated from Patients in Intensive Care Units.
Taeksoo KIM ; Jongyoun YI ; Ki Ho HONG ; Jeong Su PARK ; Eui Chong KIM
The Korean Journal of Laboratory Medicine 2011;31(1):30-36
BACKGROUND: Various virulence factors and superantigens are encoded by mobile genetic elements. The relationship between clonal background and virulence factors differs in different geographic regions. We compared the distribution and relationship of spa types and virulence genes among methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from a tertiary hospital in 2000-01 and 2007-08. METHODS: In 2000-01 and 2007-08, 94 MRSA strains were collected from 3 intensive care units at a Korean tertiary hospital. We performed spa typing and multiplex PCR for 19 superantigen genes. RESULTS: Relatively frequent spa types were t037 (40.5%), t002, t601, and t2138 in 2000-01, and t2460 (43.9%), t002, t037, t601, t324, and t2139 in 2007-08. We identified 4 novel spa types, 2 of which were designated as t5076 and t5079. Superantigen profiles were closely linked to spa types. For example, sea, sek, and seq superantigen genes were mainly detected in t037 strains. CONCLUSIONS: Major spa types differed depending on study periods, and the distribution of superantigen genes correlated with spa type.
Bacterial Typing Techniques
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DNA, Bacterial/chemistry
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Genotype
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Humans
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Intensive Care Units/statistics & numerical data
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Methicillin-Resistant Staphylococcus aureus/genetics/*isolation & purification/pathogenicity
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Microbial Sensitivity Tests
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Polymerase Chain Reaction
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Staphylococcal Infections/microbiology
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Superantigens/genetics
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Virulence/genetics
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Virulence Factors/*genetics
8.Rapid Identification of Legionella Pathogenicity by Surface-Enhanced Raman Spectroscopy.
Jing LI ; Tian QIN ; Xiao Xiao JIA ; Ai Hua DENG ; Xu ZHANG ; Wen Hui FAN ; Shuai Dong HUO ; Ting Yi WEN ; Wen Jun LIU ;
Biomedical and Environmental Sciences 2015;28(6):437-444
OBJECTIVETo establish Surface-enhanced Raman Spectroscopy (SERS) can be used as a rapid and reliable method to distinguish virulent strain and mild strain of L. pneumophila.
METHODSMortality data were collected from company departments through administrative documents, death certificates, etc. Trend analyses of cancer mortality were performed on the basis of 925 cancer deaths between 2001 and 2010.
RESULTSOur results indicated that the peaks of high virulence strains reached ⋝4000. This criterion was verified by subsequent cell experiments. In addition, we also conducted SERS rapid identification on the virulence of several collected clinical strains and obtained accurate results.
CONCLUSIONThe present study indicates that the established SERS protocol can be used as a rapid and reliable method to distinguish virulent and mildly virulent strains of L. pneumophila, which can be further used in clinical samples.
Cell Line ; Citric Acid ; chemistry ; Gold ; chemistry ; Humans ; Legionella ; isolation & purification ; pathogenicity ; Nanoparticles ; chemistry ; Spectrum Analysis, Raman ; methods ; Time Factors ; Tiopronin ; chemistry ; Virulence
9.Molecular epidemiological analysis of group A Streptococci isolated from children in Chaoyang District of Beijing, 2011: emm types, virulence factor genes and erythromycin resistant genes.
Hai Bin WANG ; Yan Yan SONG ; Yuan Hai YOU ; Heng Wei WANG ; Qin Hua HAN ; Jian Hong ZHAO ; Xiao Xi ZHANG
Biomedical and Environmental Sciences 2013;26(9):782-784