1.Molecular Typing of Salmonella typhi by Random Amplified Polymorphic DNA Analysis.
Chul Hun CHANG ; Jeong Whan SHIN ; Han Chul SON ; Chul Min KIM ; Ju Hyun LEE
Korean Journal of Clinical Microbiology 1998;1(1):51-56
BACKGROUND: In the year 1996, there were some outbreaks of Salmonella typhi infection in Pusan and therefore, the incidence of S. typhi infection was markedly increased in comparison with the previous year. To differentiate the isolates epidemiologically, a random amplified polymorphic DNA(RAPD) fingerprinting method has been developed. METHODS: A total of 9 arbitrary primers were screened with S. typhi strains isolated in Pusan, 1996. This allowed selection of a panel of primers capable of detecting DNA polymorphisms among S. typhi isolates. This panel was used to examine 54 strains of S. typhi, which had been isolated in Pusan including the cases of outbreaks that was previously characterized by phage typing. RESULTS: Four single primers and one combination of two primers were selected to discriminate the S. typhi isolates. RAPD analysis resolved the 54 strains into 20 different subtypes. At least two outbreaks were found by RAPD analysis. The isolates of E1 phage type, which are the most common in Korea, were perfectly differentiated with each other, except the strains isolated within the outbreaks. CONCLUSION: The RAPD approach is the useful epidemiologic tool to S. typhi subtyping, which is providing high discriminatory power. There were at least two outbreaks when the epidemic Salmonella infections of Pusan in 1996 had been occurred. The primers or their comb ination capable to discriminate the S. typhi isolates were described.
Animals
;
Bacteriophage Typing
;
Bacteriophages
;
Busan
;
Comb and Wattles
;
Dermatoglyphics
;
Disease Outbreaks
;
DNA*
;
Incidence
;
Korea
;
Molecular Typing*
;
Salmonella Infections
;
Salmonella typhi*
;
Salmonella*
2.Phage Typing of Staphylococcus intermedius Isolated from Canine Clinical Specimens.
Cheong Kyu PARK ; Sung Kuk KIM ; Young Ju LEE ; Sang Geon YEO ; Ki Seuk KIM
Journal of Bacteriology and Virology 2005;35(3):191-196
Bacteriophages were induced from staphylococcus intermedius isolates from dogs and used for a phage typing. Mitomycin C induction was performed on 60 strains of S. intermedius and all phages were reacted with the strains. Twenty-eight strains (46.7%) were found to be lysogenic. Based on host ranges, eight phages were selected. By using the eight phages, 129 strains isolated from canine clinical specimens were subjected to the phage typing at the routine test dilution (RTD) and 100xRTD. Typability of the phage set was 78.3%, yielding 40 phage patterns. The phage set was considered useful for differentiating S. intermedius strains isolated from dogs. None of 50 strains of S. aureus, 2 (3.6%) of 56 S. simulance strains, 13 (24.5%) of 53 S. chromogenes strains and 46 (28.1%) of 164 S. hyicus strains were typable by the phages at 100xRTD.
Animals
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Bacteriophage Typing*
;
Bacteriophages*
;
Dogs
;
Host Specificity
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Mitomycin
;
Staphylococcus intermedius*
;
Staphylococcus*
3.Molecular characterization of Vibrio cholerae phage-type 6b epidemic isolates from 1998 to 2001 in Sichuan province.
Dong-lei XU ; Hong-xia WANG ; Bao-wei DIAO ; Hong-lu LIU ; Li-feng XIONG ; Shou-yi GAO ; Biao KAN
Chinese Journal of Preventive Medicine 2009;43(5):409-412
OBJECTIVETo investigate the molecular characteristics of phage-type 6b isolates emerging in 1998-2001 cholera epidemics in Sichuan province.
METHODSIsolates were analyzed by phage-typing, pulsed field gel electrophoresis (PFGE) and ompW gene sequencing.
RESULTSAll phage-type 1b and 6b isolates in Sichuan province from 1998 to 2001 were toxigenic. A total of 24 patterns were identified after PFGE analysis, and one predominant pattern consisted of 13 isolates. Several 1b and 6b isolates from Sichuan and isolates of the 1b from other provinces showed the same PFGE pattern. Mutation in ompW gene was found in 6b isolates.
CONCLUSIONV.cholerae O1 6b isolates in Sichuan province from 1998 to 2001 have special genetic markers, and might genetically correlate with contemporaneous 1b isolates.
Bacterial Typing Techniques ; Bacteriophage Typing ; China ; epidemiology ; Cholera ; epidemiology ; microbiology ; DNA, Bacterial ; genetics ; Electrophoresis, Gel, Pulsed-Field ; Genes, Bacterial ; Genotype ; Vibrio cholerae ; classification ; genetics ; isolation & purification
4.Typing and Antimicrobial Susceptibilities of Methicillin Resistant Staphylococcus aureus (MRSA) Strains Isolated in a Hospital in Korea.
Hee Joo LEE ; Jin Tae SUH ; Yeong Sic KIM ; Walgang LENZ ; Gabriele BIERBAUM ; Klaus P SCHAAL
Journal of Korean Medical Science 2001;16(4):381-385
Methicillin-resistant Staphylococcus aureus (MRSA) strains may cause serious nosocomial infections, including pneumonia and septicemia. The rate of methicillin-resistance among S. aureus isolates in Korea is over 50%. In this study, 90 MRSA isolates from Kyung Hee University Hospital were characterized employing bacteriophage typing, pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility testing. Eighty percent of the strains could be phage-typed. The largest group or 40% of the strains belonged to lyso group III, followed by 32% of the isolates which produced a reaction with regional additional phages. Phage type 83A was most frequently encountered, followed by phage type D11. PFGE patterns confirmed the presence of two major clusters, which comprise the isolates belonging to lyso group III and the strains that were typable with regional additional phages. The latter group also contained a number of strains that were nontypable with bacteriophages. The resistance rates to ciprofloxacin, erythromycin, tetracycline, gentamicin and clindamycin were over 94%. Strains with intermediate resistance to vancomycin strains or resistance to mupirocin were not found. In conclusion, this study demonstrates that the results of phage typing are confirmed and supplemented by PFGE data.
Bacteriophage Typing
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Electrophoresis, Gel, Pulsed-Field
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Human
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*Methicillin Resistance
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Microbial Sensitivity Tests
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Staphylococcus aureus/*classification/drug effects
5.Epidemic by Salmonella I 4,5,12:i:- and Characteristics of Isolates in Korea.
Deog Yong LEE ; Esther LEE ; Jung Eun MIN ; Seong Han KIM ; Hee Bok OH ; Mi Sun PARK
Infection and Chemotherapy 2011;43(2):186-190
BACKGROUND: Salmonella I 4,[5],12:i:- was first reported as a new serovar by the Spanish National Reference Laboratory in 1997. Thereafter, several outbreaks caused by this serovar have been reported, indicating worldwide transmission. MATERIALS AND METHODS: Stool samples and patient data were collected from diarrhea cases in an outbreak at Daegu city in 2008. Salmonella isolates were characterized by phage typing, antibiotic resistance profile and flagella gene deletion. Deposited isolates in the EnterNet-Korea, acute diarrheal surveillance system, were also screened for this serovar. RESULTS: Isolates from diarrhea patients in the Daegu outbreak (2008) were identified as Salmonella I 4,[5],12:i:-. Screening the deposited isolates in the EnteroNet-Korea revealed that an unidentifed isolate in 2001 was the Salmonella I 4,[5], 12:i:-. CONCLUSIONS: Salmonella I 4,[5],12:i:-. was the causative pathogen of the 2008 foodborne outbreak of salmonellosis in Daegu City. Retrospective screening revealed that Salmonella I 4,[5],12:i:- was present in Korea as early as 2001.
Bacteriophage Typing
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Diarrhea
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Disease Outbreaks
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Drug Resistance, Microbial
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Flagella
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Gene Deletion
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Humans
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Korea
;
Mass Screening
;
Retrospective Studies
;
Salmonella
;
Salmonella Infections
6.Molecular Analysis of Salmonella enterica Serotype typhi Isolated Sporadically in Seoul City.
Hee Moo LEE ; Cheol Hyun KIM ; Jang Wook SOHN ; Mi Sun PARK ; Bok Kwon LEE ; Min Ja KIM
Korean Journal of Infectious Diseases 2000;32(5):357-365
BACKGROUND: Typhoid fever, caused by Salmonella enterica serotype Typhi, remams an important public health problem in Korea, and asymptomatic chronic carriers play a role in the endemicity. However, the molecular studies of S. typhi isolates are very limited. We characterized clinical isolates of S. typhi by molecular and phage typing tools for the extent of genetic diversity and relatedness among the isolates. METHODS: A total of 49 S. typhi isolates from sporadic cases of typhoid fever were collected in 3 university hospitals in Seoul during 1992 to 1998 and examined for in vitro susceptibility to 14 antimicrobials by disk diffusion method, ribotyping using PstI restriction enzyme, and pulsed-field gel electrophoresis (PFGE) using XbaI and Vi phage typing. The distribution of the epidemiological types and genomic DNA relatedness were analyzed. RESULTS: Forty-five out of 49 isolates were susceptible to all drugs tested. Thirty-two out of 47 were typable by phage typing and 56.3% possessed the phage type El or Ml. Forty-nine isolates divided into 6 different ribotypes (A to F) and 19 different PFGE types (AO through A17, BO) by ribotyping and PFGE analysis, respectively. Based on the 3 typing systems, 32 isolates divided into 17 different epidemiological types. The E1-A-A12 (phage type-ribotype-PFGE type) was most prevalent (18.8Fo) and isolated only in 1998, but distributed in various areas of isolation. The next prevalent M1-A-A1 (15.6%) was isolated from 1992 through 1998. The genetic relatedness based on PFGE analysis revealed that F (coefficient of similarity) values are 0.64 to 1.0 and 0.52 for A subtypes and BO type, respectively. CONCLUSIONS: We conclude that the circulating S. typhi strains in Seoul city show considerable genetic diversity, whereas most of them seems to be clonally related.
Bacteriophage Typing
;
Bacteriophages
;
Diffusion
;
DNA
;
Electrophoresis, Gel, Pulsed-Field
;
Genetic Variation
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Hospitals, University
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Korea
;
Public Health
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Ribotyping
;
Salmonella enterica*
;
Salmonella typhi
;
Salmonella*
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Seoul*
;
Typhoid Fever
7.Application of pulse field gel electrophoresis in molecular typing method on dealing with the outbreak of intestinal pathogen.
Xiao-meng WANG ; Alida YOUNAN ; Masuda TAKASHI
Chinese Journal of Preventive Medicine 2006;40(2):109-112
OBJECTIVETo explore and establish fast and sensitive typing diagnostic methods on intestinal pathogen.
METHODSAll 29 isolates of Shigella. sonnei and 12 strains of Escherichia coli O157:H7 (EHEC O157:H7) isolated in 1999 were carried out for typing by using pulse-field gel electrophoresis (PFGE) and phage typing.
RESULTSPFGE results of 4 outbreak incidents caused by Shigella. sonnei showed that the PFGE patterns of the strains isolated from the same incident were almost the same. It was also the same during analysing the 2 outbreak incidents caused by EHEC O157:H7, however, the PFGE patterns of the 3 individual strains of EHEC O157:H7 were different. Moreover, the phage typing method showed its fine typing ability while its being used in the analysing 12 strains of EHEC O157:H7.
CONCLUSIONThe fine typing ability and reliability of PFGE was helpful for finding out the heredity background of intestinal pathogen and the infectious resource of the outbreak. And the epidemic situation might thus be effectively prevented from spreading.
Bacteriophage Typing ; methods ; Disease Outbreaks ; Electrophoresis, Gel, Pulsed-Field ; Enterobacteriaceae Infections ; epidemiology ; Escherichia coli O157 ; classification ; Humans ; Japan ; epidemiology ; Shigella sonnei ; classification
8.Molecular typing of isolates from two dysentery outbreaks in Chengdu, through pulsed field gel electrophoresis technology.
Yan-Fang MIAO ; Wei HUANG ; Jian-Ping HUANG ; Zhen-Jun LI ; Dong JIN ; Zhi-Gang CUI ; Ai-Lan ZHAO ; Na LIANG ; Yan BAI
Chinese Journal of Epidemiology 2008;29(3):282-285
OBJECTIVEUsing pulsed field gel electrophoresis (PFGE) and polymerase chain reaction (PCR) typing to analyze strains isolated from two outbreaks caused by Shigella sonnei and to trace the source of infection.
METHODSVirulence genes ipaH and ial were detected by PCR and PFGE was used to subtype the isolates. Patterns were compared, using the software BioNumerics.
RESULTSWithin the 54 isolates, all were ipaH positive with 48 as ial positive. Strains from the Chongzhou outbreak were clustered into 4 PFGE patterns, with the predominant pattern accounted for 72% of the analyzed strains. The pattern of strains isolated from the cold pork with sauce was identical to the predominant pattern. The strains from Dayi outbreak were clustered into 8 PFGE patterns and the predominant pattern accounted for 56% of the test strains.
CONCLUSIONStrains from the two outbreaks were quite different and the 'cold pork with sauce' seemed to be the major source of infection, causing the outbreak of diarrhea in Chongzhou. The sources of infection of the Dayi outbreak might be complicated whereas PFGE showed a discriminatory and reproducible laboratory tool in the epidemiologic investigation on outbreaks of diarrhea.
Bacteriophage Typing ; China ; epidemiology ; Disease Outbreaks ; Dysentery, Bacillary ; epidemiology ; microbiology ; Electrophoresis, Gel, Pulsed-Field ; Food Microbiology ; Foodborne Diseases ; epidemiology ; microbiology ; Humans ; Shigella ; classification ; isolation & purification
9.Identification and molecular study on vibrio cholerae in sea products.
Zhao-rui CHANG ; Jing ZHANG ; Duo-chun WANG ; Hao-jie ZHONG ; Jin XU ; Lu RAN ; Mao-wu WANG ; Zi-jun WANG ; Biao KAN
Chinese Journal of Preventive Medicine 2007;41(4):304-306
OBJECTIVETo investigate the serologic type, phage-biotype and toxic factor of Vibrio cholerae isolated from different sea products, analyze the relation between the Vibrio cholerae in sea products and cholera epidemiology, and provide references for forecasting cholera epidemic situation and drawing out a preventing plan.
METHODThe biotype of strains isolated was analyzed by using type and phage-biotype serological methods. The toxic gene was detected by PCR.
RESULTSThe constituent ratio of V. cholerae O139, Ogawa and Inaba were, respectively, 48.44%, 20.31% and 31.25% in 64 strains of V. cholerae. The result of phage-biotype showed that the 26 strains of V. cholerae O1 were all non-epidemic strains. The result of toxic gene detecting showed that positive rate of V. cholerae O139 was higher than those of Ogawa and Inaba.
CONCLUSIONThe positive rate of toxic gene in V. cholerae O139 was high and the V. cholerae O139 was mainly in turtle, breed aquatics water and crustacean, so these sea products were the important sectors in cholera prevention and control.
Animals ; Bacteriophage Typing ; DNA, Bacterial ; genetics ; Seafood ; microbiology ; Serotyping ; Vibrio cholerae ; classification ; genetics ; isolation & purification ; Vibrio cholerae O1 ; isolation & purification ; Vibrio cholerae O139 ; isolation & purification
10.An etiological survey on a foodborne disease epidemic outbreak caused by Salmonella enteritidis.
Jin-chuan YANG ; Yang XIA ; Hui GUO ; Jing-jing XU ; Lu-mei WANG ; Jing TONG ; Lei ZHANG ; Jun-rong LIANG ; Huai-qi JING ; Zhen-jun LI
Chinese Journal of Preventive Medicine 2013;47(2):160-163
OBJECTIVETo conduct an etiological molecular epidemiological survey and laboratory test on a foodborne disease epidemic outbreak to make clear of the cause and implement effective prevention and control on it.
METHODSOn May 12th 2012, 135 kindergarten children were sent to Xuzhou City People's Hospital and Children's Hospital with gastrointestinal infection disease. A total of 34 anus swab samples and 4 vomit samples were collected from the patients. Real-time PCR rapid detection, strains separation and cultivation, phage lysis experiments, ATB automated identification system were used to make etiological detection and identification. The genomic DNA of salmonella enteritidis were typed with the pulsed-field gel electrophoresis (PFGE), cluster analysis were carried out together with the patterns of local Salmonella infections.
RESULTSChildren in 20 classes were suffered from the gastrointestinal infection among the 21 classes. There were no significant aggregation of class distribution. Among the 135 patients, 76 were boys (56.3%) and 59 were girls (43.7%). The main symptoms were fever (above 38°C), diarrhea and bellyache. Through real-time PCR detection and strains separation, 19 salmonella enteritidis were isolated from 34 anus swab samples of suspected cases and the detection rate was 56%. There were no strains detected from vomit samples. All of the 19 salmonella enteritidis showed the same serological subtype, biochemical reaction, drug sensitivity and phage lysis pattern. The salmonella enteritidis had the identical PFGE pattern (100% similarity), and were different from the pattern of local sporadic infection cases.
CONCLUSIONIt was confirmed that this was an epidemic outbreak of foodborne disease caused by homologous salmonella enteritidis by epidemiological survey, clinical information, lab etiological test and molecular typing.
Bacteriophage Typing ; Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Electrophoresis, Gel, Pulsed-Field ; Female ; Humans ; Male ; Molecular Epidemiology ; Salmonella Food Poisoning ; epidemiology ; microbiology ; Salmonella enteritidis ; classification ; isolation & purification