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
;
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
;
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
;
Korea
;
Mass Screening
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Retrospective Studies
;
Salmonella
;
Salmonella Infections
6.Detection of Antibiotic Resistant Genes in Salmonella enterica Serovar Typhimurium Isolated from Foodborne Patients in Seoul Using Multiplex-PCR.
Young Hee OH ; Mi Ok SONG ; Moo Sang KIM ; Seog Gee PARK ; Young Ki LEE
Journal of Bacteriology and Virology 2005;35(3):183-190
The frequency of antibiotic resistance among Salmonella enterica serovar Typhimurium has increased due to the transfer of multiple resistance factors. We detected the 13 antibiotic resistance genes by multiplex-PCR and compared with the results of phage typing and antibiotic disk diffusion for 49 S. typhimurium isolated from food-poisoning outbreaks in Seoul from 1999 to 2002. Resistance genes for tetracycline, streptomycin, ampicillin, sulfonamide, amino-glycoside-modifying enzyme, chloramphenicol, kanamycin, and trimethoprim were detected in 67.3%, 57.1%, 26.5%, 8.1%, 8.1%, 5%, 2.0%, and 0% of isolates, respectively. Overall 28 isolates (57.1%) possessed two or more antibiotic resistance genes. Class 1 integron carrying multidrug resistace genes, ant(3")-IaB, blaPSE, qacE delta1/sul, and tet G were amplified especially in only DT104 isolates. Among the related resistance genes for same antibiotics, strA and strB for streptomycin resistance were simultaneously detected but tetA and tetB for tetracycline were sporadically detected. DT 104 isolates contained only aadA2 and tetG.
Ampicillin
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Anti-Bacterial Agents
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Bacteriophage Typing
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Chloramphenicol
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Diffusion
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Disease Outbreaks
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Drug Resistance, Microbial
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Humans
;
Integrons
;
Kanamycin
;
R Factors
;
Salmonella enterica*
;
Salmonella*
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Seoul*
;
Streptomycin
;
Tetracycline
;
Trimethoprim
7.Molecular Typing of Cryptococcus neoformans Isolated from Korean Patients.
So Hae PARK ; Sung Il CHO ; Soo Myung HWANG
Journal of Bacteriology and Virology 2013;43(2):111-119
Cyptococcosis is generally caused by Cryptococcus neoformans, the opportunistic agent which has two species such as C. neoformans and C. gattii. Both C. neoformans and C. gattii species contain a number of genetically diverse subgroups that can be differentiated by various molecular typing methods. We conducted a molecular epidemiological analysis of 30 clinical isolates of the C. neoformans from cryptococcosis patients who had been hospitalized between 2008 and 2010 in medical centers located in Seoul and Busan in Korea. To determine the genetic diversity, 30 strains of C. neoformans were typed using PCR fingerprinting with the microsatellite specific primer of the phage M13 and the restriction fragment length polymorphism (RFLP) of orotidine monophosphosphate pyrophosphorylase (URA5) gene. All isolates were identified as serotype A, mating type MATa and molecular type VNI. The random amplified polymorphic DNA (RAPD) profiles obtained by using two primers revealed a single pattern. Our study shows that 30 strains of clinical C. neoformans are genetically homogeneous, with all of the isolates were molecular type VN1, serotype A, mating type MATa.
Bacteriophage M13
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Cryptococcosis
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Cryptococcus
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Cryptococcus neoformans
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Dermatoglyphics
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DNA
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Genetic Variation
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Humans
;
Korea
;
Microsatellite Repeats
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Molecular Typing
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Polymerase Chain Reaction
;
Polymorphism, Restriction Fragment Length
;
Uridine
8.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
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Bacteriophages
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Diffusion
;
DNA
;
Electrophoresis, Gel, Pulsed-Field
;
Genetic Variation
;
Hospitals, University
;
Korea
;
Public Health
;
Ribotyping
;
Salmonella enterica*
;
Salmonella typhi
;
Salmonella*
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Seoul*
;
Typhoid Fever
9.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
10.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