• Journal of Microbiology and Biotechnology
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• v.28 no.2
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• pp.338-346
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• 2018

Two molecular epidemiologic methods, IS6110 restriction fragment length polymorphism (IS6110-RFLP) and 24-locus mycobacterial interspersed repetitive unit-variable-number tandem repeat (MIRU-VNTR), are used worldwide in studies of Mycobacterium tuberculosis (MTB). Conversely, because of its poor resolution, pulsed-field gel electrophoresis (PFGE) is not widely used for MTB. In this study, we improved the 24-locus MIRU-VNTR and PFGE protocols and compared the effectiveness of these approaches for the molecular typing of MTB using 75 clinical isolates obtained from a cohort investigation of high-risk populations infected with MTB. The 24-locus MIRU-VNTR method demonstrated superior discriminatory ability, followed by PFGE and IS6110-RFLP. Next, we analyzed six isolates with clear epidemiologic connections; that is, isolates from patients who attended the same school. IS6110-RFLP and PFGE identified these samples as the same type. By contrast, according to MIRU-VNTR, two isolates differed from four other isolates at one locus each; one isolate was identified as Mtub29 and the other as QUB-26. In summary, the 24-locus MIRU-VNTR assay was the most useful molecular typing method among the three methods investigated due to its discriminatory power, short time required, and availability as an epidemiologic investigation tool. PFGE was the second-best method. Compared with the other loci assessed in the 24-locus MIRU-VNTR assay, the Mtub29 and QUB-26 loci appeared to exhibit greater variability during transmission.

• Journal of Microbiology
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• v.43 no.6
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• pp.546-554
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• 2005

We assessed the genetic relations and epidemiological links among bloodstream isolates of Candida albicans, which were obtained from a university hospital over a period of five years. The 54 bloodstream isolates from the 38 patients yielded 14 different karyotypes, 29 different patterns after digestion with SfiI (REAG-S), and 31 different patterns after digestion with BssHII (REAG-B) when analyzed using three different pulsed-field gel electrophoresis (PFGE) typing methods. In 11 patients with serial blood stream isolates, all strains from each patient had the same PFGE pattern. The dendrograms for all of the strains revealed that the distribution of similarity values ranged from 0.70 to 1.0 in the REAG-S patterns, and from 0.35 to 1.0 in the REAG-B patterns. Overall, the combination of the three different PFGE methods identified 31 distinct types, reflecting the results obtained using the REAG-B alone different. different Five PFGE types were shared among 22 isolates from 12 patients. These types of strains were more frequently associated with central venous catheter-related fungemia than the other 26 type strains $(92\%\;versus\;31\%;\;P<0.005)$. Of five PFGE types, four isolates were determined to be epidemiologically related: each of these types was primarily from two or three patients who had been hospitalized concurrently within the same intensive care unit. Our results suggest that the REAG-B constitutes perhaps the most useful PFGE method for investigating C. albicans candidemia and also shows that a relatively high proportion of C. albicans candidemia may be associated with exogenous acquisition of clonal strains.

• Journal of Microbiology
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• v.41 no.1
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• pp.1-6
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• 2003

The genetic relatedness of multidrug-resistant pneumococcal isolates of serotypes 19F and 23F was investigated. The DNA fragments digested with Sma I were resolved by pulsed-field gel electrophoresis (PFGE). PFGE analysis of 365. pneumoniae isolates showed 13 different patterns. Among 22 isolates of serotype 19F, 9 different PFGE patterns were present and 14 isolates of serotype 23F isolates represented 5 distinct PFGE patterns. Two isolates of serotype 19F and six isolates of serotype 23F shared the same PFGE pattern (Pattern I). Based on the genetic relatedness within the strains (one genetic cluster was defined as having more than 85% homology), we divided the pneumococcal strains into genefic clusters (Ⅰ, II, III, IV, V, and VI). The 22 strains of serotype 19F belonged to five distinct genetic clusters (I, II, III, IV, V and VI) and 14 strains of serotype 23F represented two genetic clusters (I and II ). These results showed that strains of serotype 19F are genetically more diverse than those of serotype 23F, Serotype 19F isolates with PFGE patterns H and I appeared to be less related to those of the remaining PFCE patterns (A to G) (less than 60% genetic relatedness), but those strains were genetically closely related with serotype 23f. These results suggest that the latter isolates originated from horizontal transfer of the capsular type 19F gene locus to 23F pneumococcal genotypes. In conclusion, the multidrug-resistant pneumococcal isolates of serotype 19f and 23F isolated in Korea are the result of the spread of a limited number of resistant clones.

• Journal of Life Science
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• v.16 no.4
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• pp.664-671
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• 2006

We obtained 424 Salmonella enterica serovar Typhi isolates from sporadic cases of infection in Busan during 1996 to 2005. We investigated the trend of antimicrobial resistance and molecular typing by pulsed-field gel electrophoresis (PFGE). Of the total 424 isolates, 6 strains (1.4%) were multidrug-resistant (MDR) S. enterica serovar Typhi isolates, 2 strains (0.5%) were resistant to only nalidixic acid, and the remaining 416 strains (98.1%) were fully susceptible to the 18 antimicrobial agent. PFGE of XbaI-digested chromosomal DNA was performed on 50 sporadic S. enterica serovar Typhi isolates with the objective of investigating the extent of genetic diversity of these isolates in our region. We could find that these isolates were much more heterogeneous and at least 32 different PFGE patterns were generated according by dice coefficient, between 0.69 and 1.0. Restriction fragment patterns consisted of 13 to 18 fragments ranged in size from 20 to 630 kb. The results confirmed that PFGE would be an useful tool for investigating surveillance of sporadic or outbreak case and assessing clonality for S. enterica serovar Typhi in Busan area. Our finding will be valuable in developing rational strategies to control this pathogen and setting the basis of an effective PulseNet system in Korea.

• Journal of Environmental Health Sciences
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• v.34 no.2
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• pp.170-174
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• 2008

The strains of Salmonella othmarschen and Norovirus isolated from outbreak A, the funeral ceremonies incidents in Ouri-shi of Oyeonggido in, 2007, were subjected to genotyping by using PFGE and extraction of RNA by using RT-PCR. 24 Salmonella othmarschen were isolated from 37 cases (patients 12 cases, cookers 4 cases, foods 7cases, knife, kitchen board, dish towel 4 cases, etc) and PFGE patterns showed that 22 strains isolated from the same incident were almost of the same pattern and 2 strains exhibited 50% of the pattern. Antimicrobial susceptibility patterns were such that there were 22 case susceptibility patterns and 2 case resistance patterns (strain 4: AM, CF, CIP, NA, TE TIC and strain 13: CF, CZ). Norovirus G1 (1case), Norovirus G2 (7cases), Norovirus G1,G2 (1case) were detected by using RT-PCR. The fine typing ability and reliability of PFGE and BioNumerics are helpful for establishing out the foodborne pathogens of outbreaks.

• Korean Journal of Veterinary Service
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• v.32 no.2
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• pp.155-163
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• 2009

Fowl typhoid (FT) is a septicemic disease caused by Salmonella Gallinarum. The purpose of this study was to investigate the antimicrobial resistance and pulsed-field gel electrophoresis (PFGE) patterns of S. Gallinarum isolated from chicken. During 1999 to 2004, there was isolated a total of 100 strains in liver and spleen. The biochemical characteristics of S. Gallinarum isolates was nonmotile, no production of H$_2$S, glucose gas, non-fermented rhamnose, indole-negative, fermentation of dulcitol, mannitol, maltose, and ornithine decarboxylase. At antimicrobial susceptibility, all of isolates were susceptible to amoxicillin/clavulanic acid, amikacin, neomycin, kanamycin, norfloxacin and enrofloxacin. One hundred isolates were divided into 54 resistant patterns and 37 strains was 6-multi drug resistance. PFGE of Xba I restriction fragments of S. Gallinarum isolates was 20 patterns.

• Biomedical Science Letters
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• v.11 no.3
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• pp.289-293
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• 2005

Two housekeeping genes, of Listeria monocytogenes dat and hlyA, were analyzed in a set of 28 isolates from different sources to estimate their genetic diversities. These strains were previously characterized by pulsed-field gel electrophoresis. Complete gene sequences for dat (465 bp) and hlyA (584 bp) had sequence similarity of $99.87-100\%$ S and $99.96-100\%$ S among isolates, respectively. Also, we found that the numbers of sequence types (ST) were about 3-fold less than those of PFGE types (3 STs versus 11 PFGE types). There was, however, a good correlation between the PFGE patterns and phylogenetic grouping of two gene sequences among the isolates. Further studies on analyzing additional loci would increase the discriminatory power of sequence typing for L. monocytogenes strains.

• Korean Journal of Veterinary Research
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• v.50 no.3
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• pp.239-246
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• 2010

Among 203 avian pathogenic Escherichia coli (APEC) isolated from poultry with colibacillosis in korea, 14 isolates were selected from total 68 isolates transferred R plasmid and classified into 5 groups on the basis of antimicrobial minimal inhibitory concentration (MIC) pattern, farm source and O serotype. An association between clonal origin and R plasmid of them was investigated by R plasmid profile, restriction endonuclease analysis and pulsed-field gel electrophoresis (PFGE). The strains that showed the same or very similar antimicrobial MIC pattern, but different farm source and O serotype, revealed different PFGE pattern, which seemed to be different clonal origin. And the strains that showed the same MIC pattern and O serotype, revealed different PFGE pattern, seemed to be originated from different clone. Also the strains showing the same MIC pattern and farm source, but different O serotype, revealed to be different clonal origin. The strains that showed the same or similar MIC pattern, farm source, and O serotype, revealed identical or similar PFGE pattern, which seemed to belong to be one clone. Meanwhile, horizontal transfer of R plasmid seems to be common in APEC with regardless of O serotype and clone of the strains. These results indicate that rapid and accurate epidemiological survey of APEC can be possible by the combination of O serotyping, plasmid profiling and PFGE analysis following the classification of them into groups of antimicrobial drug resistance pattern.

• Journal of Life Science
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• v.15 no.2 s.69
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• pp.161-168
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• 2005

In this study, we did the molecular typing of 39 environmental Legionella pneumophila serogroup 1 isolates collected from 2001-2003 in Busan using the pulsed-filed gel electrophoresis (PFGE). PFGE of SfiI fragments were divided into 10 pulsotypes $(A\~J)$, corresponding to $<65\%$ similarity and a subtype within each pulsotype was characterized by $>84\%$ similarity. The major cluster was pulsotype E $(46.2\%)$, which included 18 isolates and was divided into 4 subtypes $(E1\~E4)$. PFGE of NotI fragments were divided into 8 pulsotypes $(a\~h)$, corresponding to $<60\%$ similarity and a subtype within each pulsotype was characterized by $100\%$ similarity. The major cluster was pulsotype f $(38.5\%)$, which included 15 isolates. The ATCC type strain L. pneumophila serogroup 1 was identified as a different molecular pulsotype compare to the Busan isolates. It is possible that L. pneumophila serogroup 1 isolated in Busan with specific DNA pattern is comparable with those isolation in other cities in Korea.

• Korean Journal of Microbiology
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• v.53 no.3
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• pp.156-162
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• 2017

Legionnaires' disease (LD) is a severe and potentially fatal pneumonia caused by colonization of human-made water system and subsequent aerosolization and inhalation of Legionella bacteria. A total of 147 Legionella strains was isolated from environmental water sources from public facilities in Gyeonggi-do, South Korea. The distribution of Legionella isolates was investigated according to facility type, and sample type. L. pneumophila was distributed broadly throughout Gyeonggi-do, accounting for 85.7% of the isolates, and L. pneumophila serogroup (sg) 1 predominated in all of the public facilities. L. wadsworthii predominated among non-L. pneumophila species. We performed comparative analyses of L. pneumophila sg 1 isolated from environment water of public facilities in Gyeonggi-do by pulsed field gel electrophoresis (PFGE) and sequence-based typing (SBT). Thirty-two isolates were classified into 22 types by PFGE and 9 sequence types (STs) by SBT and categorized into 3 groups. ST1 was the most prevalent sequence type and two STs obtained in this study had unique allelic profiles. The use of SBT data from different countries for epidemiology study of LD constitutes a technically uncomplicated and relatively easy method for strain subtyping, especially compared to other contemporary techniques.