1.Comparison of Nontoxigenic and Neurotoxigenic Clostridium butyicum by Molecular Typing Methods.
Xingmin WANG ; Tad Ahiro KARASAWA ; Tsuneo MAEGAWA ; Shunji KOZAKI ; Kentaro TSUKAMOTO ; Shinichi NAKAMURA
Journal of the Korean Society for Microbiology 2000;35(5):369-369
No Abstract Available.
Clostridium*
;
Molecular Typing*
4.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
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Bacteriophage Typing
;
Bacteriophages
;
Busan
;
Comb and Wattles
;
Dermatoglyphics
;
Disease Outbreaks
;
DNA*
;
Incidence
;
Korea
;
Molecular Typing*
;
Salmonella Infections
;
Salmonella typhi*
;
Salmonella*
5.Molecular typing of 12 Brucella strains isolated in Guizhou province in 2010-2013.
Yue WANG ; Hong CHEN ; Ying LIU ; Jingzhu ZHOU ; Shijun LI ; Email: ZJUMEDJUN@163.COM. ; Yan HANG ; Guangpeng TANG ; Dingming WANG ; Guichun CHEN
Chinese Journal of Preventive Medicine 2015;49(9):788-791
OBJECTIVETo identify and characterize the Brucella strains from Guizhou province in 2010-2013.
METHODSA total of 12 strains of Brucella suspicious bacteria were isolated in Guizhou province from 2010 to 2013. Four strains (GZLL3, GZLL4, GZLL11 and SH2) were isolated from goat blood samples and eight strains (SH4, GZZY, GZSQ, GZZA, BR13001, BR13004, BR13005 and BR13006) were isolated from blood samples of patient 12 Brucella suspicious strains were identified and characterized using conventional methods. Brucella genus specific gene BCSP31-based PCR (BCSP31-PCR) was used to identify the genus of Brucella and IS711 insert sequence-based PCR (AMOS-PCR) was applied to identify the species of Brucella strains. Goats and patients originated Brucella strains were comparatively analysed using Pulse-field Gel Electrophoresis (PFGE).
RESULTSBoth of conventional methods and PCR identified the 12 Brucella suspicious strains as B. melitensis biotype 3. BCSP31-PCR identification results showed that a specific DNA bands (223 bp) were detected in all the 12 strains and positive control samples with no DNA band in negative samples. AMOS-PCR amplified a 731 bp-DNA bands in all the 12 strains, with 731 bp, 498 bp and 275 bp in M5, S2 and A19 strains, respectively, and no DNA band was detected in the negative control samples. PFGE analysis showed that 12 Brucella isolates from patients and goats showed consistent PFGE patterns with the digestion of restriction enzyme Xba I.
CONCLUSIONThe epidemic species/type of Brucella in both human and animal in Guizhou province was B. melitensis biotype 3 and goat was the main animal source of infection of brucellosis in Guizhou province.
Animals ; Bacterial Typing Techniques ; Brucella ; classification ; Brucellosis ; epidemiology ; China ; epidemiology ; DNA, Bacterial ; Goats ; Humans ; Molecular Typing ; Polymerase Chain Reaction
6.Molecular Biology of Cutaneous Fungal Infection.
Hanyang Medical Reviews 2006;26(4):52-58
Traditional identification of fungi is based on their gross morphology and microscopic findings. It takes a long time and needs experience and sometimes it is difficult because their morphology may change. Nucleic acid detection methods such as PCR have become a common tool for microbial identification and diagnosis. Molecular typing techniques provide information to detect their infection routes. They are fast, sensitive, and reproducible. Pure DNA, the targets, and methods are important for good results. There are lots of techniques like sequencing, RFLP, RAPD, and hybridization. Each method has its advantage and limitation, and proper selection is essential. In clinical application, careful consideration will reduce errors. There is a limited number of reports about cutaneous fungal infection. We review the molecular methods for fungal identification and typing, and discuss the advantages and limitations of the methods.
Diagnosis
;
DNA
;
DNA, Ribosomal
;
Fungi
;
Molecular Biology*
;
Molecular Typing
;
Polymerase Chain Reaction
;
Polymorphism, Restriction Fragment Length
8.Molecular Classification of Colorectal Cancers and Clinical Application.
So Yeon JEON ; Won Kyu KIM ; Hoguen KIM
The Korean Journal of Gastroenterology 2016;68(6):297-302
The molecular genetics of colorectal cancers (CRCs) is among the best understood of common human cancers. It is difficult to predict the prognosis and/or to predict chemoresponding in CRC patients. At present, prognosis is based predominantly on the tumor stage and pathological examination of the disease. Molecular classification of CRCs, based on genomics and transcriptomics, proposed that CRCs can be classified into at least three-to-six subtypes, depending on the gene expression pattern, and groups of marker genes representing to each subtype have also been reported. Gene expression-based subtyping is now widely accepted as a relevant source of disease stratification. We reviewed the previous studies on CRC subtyping, international consortium dedicated to large-scale data sharing and analytics recently established four consensus molecular subtypes with distinguishing features. Predictive markers identified in these studies are under investigation and large-scale clinical evaluations of molecular markers are currently in progress.
Classification*
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Colonic Neoplasms
;
Colorectal Neoplasms*
;
Consensus
;
Gene Expression
;
Genomics
;
Humans
;
Information Dissemination
;
Molecular Biology
;
Molecular Medicine
;
Molecular Typing
;
Prognosis
9.Comparison of Pulsed-field Gel Electrophoresis, Amplified Fragment Length Polymorphism, and Infrequent Restriction Site-Polymerase Chain Reaction for Molecular Typing of Escherichia coli and Staphylococcus aureus Strains.
Sang Il KIM ; Jin Hong YOO ; Yoon Kyung CHO ; Dong Gun LEE ; Seong Heon WIE ; Jung Hyun CHOI ; Yang Ree KIM ; Wan Shik SHIN ; Moon Won KANG
Korean Journal of Infectious Diseases 1999;31(6):474-480
BACKGROUND: Pulsed-field gel electrophoresis (PFGE) has been regarded a standard method for genotyping in epidemiologic studies. However, it is tedious and time-consuming to perform. Two alternative genotyping methods have recently been developed using the polymerase chain reaction (PCR):amplified fragment length polymorphism (AFLP) and infrequent restriction site-polymerase chain reaction (IRS-PCR). These methods have not yet been applied yet to common pathogens such as Staphylococcus aureus. The purpose of this study was to determine the applicability of AFLP and IRS-PCR for the genotyping of E. coli and S. aureus isolates. METHODS: We performed PFGE, AFLP, and IRS-PCR on clinical isolates of E. coli (n=27) and S. aureus (n=30). We assessed each method in terms of discriminatory power, quality, and efficiency. RESULTS: In E. coli, the discriminatory powers of IRS-PCR and AFLP were comparable to that of PFGE. PFGE discerned 24 (88.8%) out of 27 strains, IRS-PCR discerned 22 (81.5%) out of 27, and AFLP discerned 25 (92.6%) out of 27. In the case of S. aureus, PFGE discerned 27 (90%) out of 30 strains, while both IRA-PCR and AFLP discerned 12 (40%) out of 30. The test-ing took four days to complete with PFGE, two days with AFLP, and was completed within one day with IRS-PCR. IRS-PCR showed better resolution than both PFGE and AFLP. CONCLUSION: In cases of E. coli, AFLP and IRS-PCR could be good alternatives for epidemiologic typing, as they offer better efficiency and comparable discriminatory power to PFGE. On the other hand, IRS-PCR and AFLP do not seem to be suitable for the strain-to-strain differentiation of S. aureus.
Electrophoresis, Gel, Pulsed-Field*
;
Escherichia coli*
;
Escherichia*
;
Hand
;
Molecular Typing*
;
Polymerase Chain Reaction
;
Staphylococcus aureus*
;
Staphylococcus*
10.Plasmid DNA and Random Amplified Polymorphic DNA (RAPD) Analyses of Vancomycin-Resistant Enterococcus faecium Isolates.
Won Pyo HONG ; Min KIM ; Jeong Won SONG ; Soo Hyun KIM ; Jong Hee SHIN ; Soon Pal SUH ; Dong Wook RYANG
Korean Journal of Clinical Pathology 1998;18(3):379-385
BACKGROUND: Vancomycin-resistant enterococci (VRE) have been increasingly reported worldwide. The understanding of VRE dissemination in the hospital requires a molecular typing of the strains. VRE appeared recently in Chonnam University Hospital. The purpose of this study is to analyse the strains for their genetic relatedness. METHODS: Nine vancomycin-resistant E. faecium isolates, collected from six patients during 1995-1996 in Chonnam University Hospital, were typed using plasmid DNA and RAPD analyses. The plasmid DNA of the isolates was obtained by a alkaline lysis method. For RAPD, eight random primers were used. The cluster analysis was performed by NTSYS-pc (numerical taxonomy system and multivariate analysis system, version 1.50, Applied Biostatistics Inc., CA). RESULTS: Nine VRE isolates were separated into two different molecular types (group A and B) by the plasmid DNA patterns, which were agreed with the RAPD results: the isolates of each group showed the same plasmid DNA patterns and high similarity values in the RAPD analysis. Group A was consisted of two strains isolated from two patients who were admitted at the same room in May 1995. Seven strains of group B were isolated from four patients in the different wards during June 1995 to June 1996. CONCLUSIONS: Nine VRE isolates from six patients were typed to two groups by plasmid DNA or RAPD analysis. These results suggested the intrahospital spread of two clonal strains of vancomycin-resistant E. faecium.
Biostatistics
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Classification
;
DNA*
;
Enterococcus faecium*
;
Enterococcus*
;
Humans
;
Jeollanam-do
;
Molecular Typing
;
Multivariate Analysis
;
Plasmids*