• 한국동물위생학회지
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• 제32권3호
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• pp.257-264
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• 2009

Subtyping of Brucella abortus isolates is epidemiologically important for monitoring of bovine brucellosis outbreaks. Pulsed-field gel electrophoresis (PFGE) is considered as a gold standard of molecular typing methods to study the DNA polymorphisms of bacteria. In this study, we analyzed using PFGE the DNA fragment profiles of B. abortus isolated in Gyeongbuk province from 1998 to 2006. The genomic DNA was digested with the restriction endonuclease Xba I, Xho I and Smi I followed gel electrophoresis. No distinguishable patterns of the genomic DNA digested with Xba I and Xho I were observed among the field isolates of B. abortus tested in this study. But Smi I restriction enzyme resulted in two PFGE patterns consisting of 13-15 bands that ranged in size from 33 to 668bp by standard marker. The cluster analysis by DNA fingerprinting software showed 93.75% similarity between two PFGE patterns. No different PFGE patterns were recognized among the isolates originated from various years, regions and cow breeds.

• 생명과학회지
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• 제16권4호
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• pp.664-671
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• 2006

1996년부터 2005년까지 부산지역에서 분리된 Salmonella enterica serovar Typhi 균주에 대한 항균제 내성 변화양상 및 Pulsed-field gel electrophoresis(PFGE)를 이용한 분리주의 분자역학적 형별을 분석하였다. 전체 424주에 대한 항균제 감수성 시험결과 multidrug-resistant (MDR) 6주(1.4%)와 nalidixic acid에만 내성을 보이는 2주를 제외한 나머지 416주(98.1%)가 시험 항균제 18종 모두에 감수성을 보였다. 부산지역 분리 장티푸스균의 유전적 이질성을 확인하고자 실시한 산발 분리 50주의 PFGE/XbaI 시험결과, 최소 32종의 다양한 패턴이 나타났다. 각 패턴별로 제한효소 절편 수는 13개에서 18개까지였고, 절편크기는 약 20 kb에서 630 kb 범위였다. 본 시험결과 부산지역의 장티푸스의 산발 또는 집단 발생시 PFGE는 유용한 역학적 지표로 사용가능함을 알 수 있었으며 또한 전국적 PulseNet 구축의 기초 자료로서 활용도가 높을 것으로 사료된다.

• 대한임상검사과학회지
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• 제39권3호
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• pp.183-189
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• 2007

Shigellosis is the most common bacterial gastroenteritis both in developing and developed countries, but bacteremia due to Shigella spp. is very rare. In developed countries recent shigellosis is mostly caused by S. sonnei, but S. flexeri infection is rare. We had rare cases of S. flexeri infections in a family in the Jeonbuk Province: an 8-year-old boy with bacteremic shigellosis and 10- and 12-year-old brothers with diarrhea. The isolates had identical biochemical characteristics, and were resistant to ampicillin, chloramphenicol, and co-trimoxazole. PFGE pattern of Not I-restricted genomic DNA suggested that the isolate from blood was closely related to the two strains isolated from stool which had an identical PFGE pattern.

• 미생물학회지
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• 제35권3호
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• pp.197-204
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• 1999

1996년 경기도, 강원도 충청북도, 전라남도, 전라북도 등 일부지역에서 채집된 들쥐들로부터 분리된 22주의 렙토스피라균의 단클론 항체에 대한 반응양상 및 genomic DNA 의 pulsed-field gel electrophoresis pattern을 분석하였다. 혈청군 Icterohemorrhagiae 내의 균주로 면역하여 제조한 7가지 단클론 항체에 대한 분리균주들의 반응은 모두 혈청형 lai 와 같은 pattern을 보였다. Not I 제한효소 절단 DNA 의 PFGE에서 JR34, JR57, JR77, JT82, JR86, JR109, NR4, NR6, NR13, CR3, KR48, NR2, NR8, NR9, NR10, NR11, NR12, JR58, 및 JR62 주 들은 모두 혈청형 lai 와 유사한 profile을 보였으며 940 kb와 63kb 사이에 13개의 절편 band를 보였다. JR89주는 혈청형 lai 및 다른 분리균주에서 관찰되지 않은 1000 kb band 와 460 kb band 가 관찰되었다.표준균주 혈청형 lai, birkini, gem, mwogolo, canicola 등은 각기 완전히 다른 pattern을 보였으며 혈청형 yeonchon 은 lai 와 같은 pattern을 보였다. UPGMA방법에 의한 dendrogram 분석결과 분리주는 lai 및 yeonchon 혈청형과 81~85%, 기타 혈청형과는 62% 이하의 유사도를 나타내어 항원분석법에 의한 혈청형 동정결과와 일치하였다. Asc I 제한요소 절단 DNA 의 PFGE에서는 JR34, JR77, JR82, JR109, NR6, NR13, CR3, KR48, 및 JR57, JR58, JR62, JR86, NR2, NR3, NR4, NR8, NR9, NR10, NR11, NR12 주들은 모두 1900 kb 에서부터 380kb 사이에 3개의 절편 band를 보였으며 이는 표준균주 lai 와 같은 pattern 이었다. JR89주는 다른 분리균주와는 달리 1640 kb 대신 650kb 의 band를 보였다. Fse I 제한효소 절단 PFGE 에서는 JR57, JR77, JR82, JR86, JR109, NR4, NR6, NR13, CR3, KR48주 및 JR34, NR2, NR3, NR9, NR10 주들은 모두 1900 kb 와 280kb 사이에 5개의 절편 band를 보였으며 이는 표준균주 lai 및 yeonchon 과 같은 pattern 이었다. 그러나 JR89주에서는 280kb 가 나타나지 않아 다른 분리균주와 구분되었다.

• Journal of Microbiology
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• 제43권6호
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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.

• 대한수의학회지
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• 제50권3호
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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 Microbiology and Biotechnology
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• 제10권3호
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• pp.405-409
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• 2000

We have studied the genome of an obligately commensal thermophile, Symbiobacterium toebii. The chromosome was extracted from pure cultures of S. toebii recently established. Total DNA of S. toebii was resolved by pulsed-field gel electrophoresis (PFGE) into discrete numbers of fragments by digenstion with the endonuclease SspI, SpeI, XbaI, and HpaI. Estimated sizes of fragments produced by the four enzymes and their sum consistently yielded a total genome size of 2.8 Mb. Because restriction endonucleases NotI and SwaI, recognizing 8 bp, released too many fragments, these enzymes could not be used for the estimation of the genome size. Considering no mobility of undigested genome under PFGE, the genome of S. toebii appears to be circular. The presence of extrachromosomal DNA in S. toebii was excluded by the results of the conventional 1% agarose gel electrophoresis and the field inversion gel electrophoresis of undigested S. toebii DNA.

• 한국동물위생학회지
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• 제30권3호
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• pp.353-362
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• 2007

A total of 1,354 samples was collected from bovine and porcine carcass from January 2005 to December 2006 in a slaughter house. Twenty five strains(1.8%) of Listeria monocytogenes were isolated from 1,354 samples using selective media. Ten(1.4%) L monocytogenes were isolated from the 677 of bovine carcasses, and 15(2.2%) were isolated from the 677 of porcine carcasses. Among 15 L mono-cytogenes from porcine, 11 siolates were serovars 1/2c, followed by 1/2b (3 strains, 20.0%) and 1/2a(1 strain) Out of 10 bovine samples, positive cases in 1/2a were 9 strains (90.0%), 1/2b were 1 strains(10.0%). PCR primers were selected to amplify a 520-base pair(bp) DNA fragment from the listeolysin O gene (hlyA) of L mono-cytogenes. A 520-bp product was detected in PCR with DNA from L monocytogenes, but not from the other Listeria species tested. A total of 25 L monocytogenes strains were analysed by PFGE after digestion with Apa I. PFGE analysis of genomic DNA showed the $14{\sim}18$ fragments ranging in size from 30 to 550 kb, resulting in 14 patterns.

• Tuberculosis and Respiratory Diseases
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• 제77권4호
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• pp.172-177
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• 2014

Background: Pseudomonas aeruginosa infection is particularly associated with progressive and ultimately chronic recurrent respiratory infections in chronic obstructive pulmonary disease, bronchiectasis, chronic destroyed lung disease, and cystic fibrosis. Its treatment is also very complex because of drug resistance and recurrence. Methods: Forty eight cultures from 18 patients with recurrent P. aeruginosa pneumonia from 1998 to 2002 were included in this study. Two or more pairs of sputum cultures were performed during 2 or more different periods of recurrences. The comparison of strains was made according to the phenotypic patterns of antibiotic resistance and chromosomal fingerprinting by pulsed field gel electrophoresis (PFGE) using the genomic DNA of P. aeruginosa from the sputum culture. Results: Phenotypic patterns of antibiotic resistance of P. aeruginosa were not correlated with their prior antibiotic exposition. Fifteen of 18 patients (83.3%) had recurrent P. aeruginosa pneumonia caused by the strains with same PFGE pattern. Conclusion: These data suggest that the most of the recurrent P. aeruginosa infections in chronic lung disease occurred due to the relapse of prior infections. Further investigations should be performed for assessing the molecular mechanisms of the persistent colonization and for determining how to eradicate clonal persistence of P. aeruginosa.

• BMB Reports
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• 제29권6호
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• pp.569-573
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• 1996

Clinical strains of Mycobacterium tuberculosis, M. terrae complex, M. gordonae, M. avium-intracellulae complex, and M. fortuitum from Korean patients were isolated and analyzed by comparing large restriction fragment (LRF) patterns produced by digestion of genomic DNA with infrequent-cutting endonucleases like AsnI and XbaI. and pulsed-field gel electrophoresis (PFGE). Three M. tuberculosis, two M. terrae complex, two M. gordonae, two M. avium-intracellulae complex, and two M. fortuitum strains were compared by using AsnI and XbaI. and this allowed easy visual separation of all epidemiologically unrelated strains. PFGE exhibits different DNA restriction patterns which are easy to compare. Genome size of the strains roughly ranged from 3020 to 3335 kb. The LRF patterns are useful for epidemiologic studies of tuberculosis with regard to drug resistance.