• 제목/요약/키워드: Multilocus sequence analysis

검색결과 59건 처리시간 0.029초

Genetic Diversity and Antibiotic Resistance of Enterococcus faecalis Isolates from Traditional Korean Fermented Soybean Foods

  • Lee, Jong-Hoon;Shin, Donghun;Lee, Bitnara;Lee, Hyundong;Lee, Inhyung;Jeong, Do-Won
    • Journal of Microbiology and Biotechnology
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    • 제27권5호
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    • pp.916-924
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    • 2017
  • Eighty-five Enterococcus faecalis isolates collected from animals (40 isolates), meju (a Korean fermented soybean product; 27 isolates), humans (10 isolates), and various environmental samples (8 isolates) were subjected to multilocus sequence typing (MLST) to identify genetic differences between samples of different origins. MLST analysis resulted in 44 sequence types (STs), and the eBURST algorithm clustered the STs into 21 clonal complexes (CCs) and 17 singletons. The predominant STs, ST695 (21.1%, 18/85) and ST694 (9.4%, 8/85), were singletons, and only contained isolates originating from meju. None of the STs in the current study belonged to CC2 or CC9, which comprise clinical isolates with high levels of antibiotic resistance. The E. faecalis isolates showed the highest rates of resistance to tetracycline (32.9%), followed by erythromycin (9.4%) and vancomycin (2.4%). All isolates from meju were sensitive to these three antibiotics. Hence, MLST uncovered genetic diversity within E. faecalis, and clustering of the STs using eBURST revealed a correlation between the genotypes and origins of the isolates.

Characterization of Pseudomonas syringae pv. syringae, Causal Agent of Citrus Blast of Mandarin in Montenegro

  • Ivanovic, Zarko;Perovic, Tatjana;Popovic, Tatjana;Blagojevic, Jovana;Trkulja, Nenad;Hrncic, Snjezana
    • The Plant Pathology Journal
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    • 제33권1호
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    • pp.21-33
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    • 2017
  • Citrus blast caused by bacterium Pseudomonas syringae is a very important disease of citrus occuring in many areas of the world, but with few data about genetic structure of the pathogen involved. Considering the above fact, this study reports genetic characterization of 43 P. syringae isolates obtained from plant tissue displaying citrus blast symptoms on mandarin (Citrus reticulata) in Montenegro, using multilocus sequence analysis of gyrB, rpoD, and gap1 gene sequences. Gene sequences from a collection of 54 reference pathotype strains of P. syringae from the Plant Associated and Environmental Microbes Database (PAMDB) was used to establish a genetic relationship with our isolates obtained from mandarin. Phylogenetic analyses of gyrB, rpoD, and gap1 gene sequences showed that P. syringae pv. syringae causes citrus blast in mandarin in Montenegro, and belongs to genomospecies 1. Genetic homogeneity of isolates suggested that the Montenegrian population might be clonal which indicates a possible common source of infection. These findings may assist in further epidemiological studies of this pathogen and for determining mandarin breeding strategies for P. syringae control.

First Report of Pectobacterium aroidearum Causing Soft Rot on Zamioculcas zamiifolia

  • Kyoung-Taek Park;Soo-Min Hong;Leonid N. Ten;Chang-Gi Back;Seung-Yeol Lee;In-Kyu Kang;Hee-Young Jung
    • 식물병연구
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    • 제29권4호
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    • pp.445-451
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    • 2023
  • Zamioculcas zamiifolia is a popular indoor ornamental plant in Korea. In August 2021, a severe outbreak of soft rot disease affected Z. zamiifolia in Emseong, Chungcheongbuk-do, Korea. Infected plants displayed wilting, water-soaked lesions, stem collapse, and green-brown discoloration. The bacterial strain KNUB-05-21 was isolated from infected stems and identified as Pectobacterium aroidearum using 16S rRNA nucleotide sequencing and multilocus sequence analysis based on partial sequences of dnaX, leuS, and recA genes. Confirmation of its affiliation with P. aroidearum was also obtained through biochemical and morphological characterization. To confirm the pathogenicity of strain KNUB-05-21, its suspension was injected into Z. zamiifolia stems. Within a week, soft rot developed on the stems, exhibiting symptoms similar to those observed in field-infected plants. The reisolated strain was identical to those of P. aroidearum. Before this study, P. aroidearum was not reported as a causative pathogen of Z. zamiifolia soft rot in Korea.

최근 4년간 대전지역에서 분리된 KPC-2 생성 Klebsiella pneumoniae의 역학적 연구 (Epidemiological Study of KPC-2 Producing Klebsiella pneumoniae Isolated in Daejeon During a 4-Year Period)

  • 조혜현
    • 대한임상검사과학회지
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    • 제54권4호
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    • pp.265-272
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    • 2022
  • Carbapenemase 생성 장내 세균(carbapenemase-producing Enterobacteriaceae, CPE) 중 특히 KPC-2 생성 Klebsiella pneumoniae의 출현과 확산은 전 세계적으로 급격히 증가하고 있으며, 심각한 공중 보건 위협이 되고 있다. 이러한 KPC-2 생성 K. pneumoniae의 역학과 특징은 지역 및 기간에 따라 다르기 때문에, 본 연구에서는 2017년 3월부터 2020년 12월까지 대전지역의 3차 병원에서 분리된 carbapenem 내성 K. pneumoniae (carbapenem-resistant K. pneumoniae, CRKP) 78 균주를 대상으로 carbapenemase 유전자를 분석하고, 이에 대한 역학 관계를 조사하였다. 항균제 감수성 양상은 디스크 확산법으로 확인하였고, carbapenemase 유전자의 분석을 위해 PCR과 DNA 염기서열분석을 수행하였다. 또한, 역학관계는 MLST (multilocus sequence typing)를 통해 조사하였다. 78 균주의 CRKP 중 35 균주(44.9%)가 carbapenemase 생성 K. pneumoniae였으며, 주요 carbapenemase 유형은 KPC-2 (30주, 85.7%)로 확인되었다. NDM-1과 NDM-5는 각각 4 균주(11.4%)와 1 균주(2.9%)에서 확인되었다. MLST 분석에서 10개의 sequence type (ST)이 확인되었고, 가장 흔한 ST는 ST307 (51.4%, 18/35)이었다. ST307 균주는 모두 KPC-2 생성 K. pneumonia였고, 다제내성으로 확인되었다. 또한, ST307은 4년 동안 지속적으로 출현하였다. 이러한 결과는 KPC-2 생성 K. pneumoniae ST307의 확산을 방지하기 위해 지속적인 모니터링과 적절한 감염 관리가 필요할 것으로 사료된다.

Infection Density Dynamics and Phylogeny of Wolbachia Associated with Coconut Hispine Beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), by Multilocus Sequence Type (MLST) Genotyping

  • Ali, Habib;Muhammad, Abrar;Hou, Youming
    • Journal of Microbiology and Biotechnology
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    • 제28권5호
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    • pp.796-808
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    • 2018
  • The intracellular bacterium Wolbachia pipientis is widespread in arthropods. Recently, possibilities of novel Wolbachia-mediated hosts, their distribution, and natural rate have been anticipated, and the coconut leaf beetle Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), which has garnered attention as a serious pest of palms, was subjected to this interrogation. By adopting Wolbachia surface protein (wsp) and multilocus sequence type (MLST) genotypic systems, we determined the Wolbachia infection density within host developmental stages, body parts, and tissues, and the results revealed that all the tested samples of B. longissima were infected with the same Wolbachia strain (wLog), suggesting complete vertical transmission. The MLST profile elucidated two new alleles (ftsZ-234 and coxA-266) that define a new sequence type (ST-483), which indicates the particular genotypic association of B. longissima and Wolbachia. The quantitative real-time PCR analysis revealed a higher infection density in the eggs and adult stage, followed by the abdomen and reproductive tissues, respectively. However, no significant differences were observed in the infection density between sexes. Moreover, the wsp and concatenated MLST alignment analysis of this study with other known Wolbachia-mediated arthropods revealed similar clustering with distinct monophyletic supergroup B. This is the first comprehensive report on the prevalence, infection dynamics, and phylogeny of the Wolbachia endosymbiont in B. longissima, which demonstrated that Wolbachia is ubiquitous across all developmental stages and distributed in the entire body of B. longissima. Understanding the Wolbachia infection dynamics would provide useful insight to build a framework for future investigations, understand its impacts on host physiology, and exploit it as a potential biocontrol agent.

Prevalence and Characteristics of Antimicrobial-Resistant Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus from Retail Meat in Korea

  • Kim, Yong Hoon;Kim, Han Sol;Kim, Seokhwan;Kim, Migyeong;Kwak, Hyo Sun
    • 한국축산식품학회지
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    • 제40권5호
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    • pp.758-771
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    • 2020
  • This study was to investigate the prevalence and characteristics of antimicrobial-resistant Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) from 4,264 retail meat samples including beef, pork, and chicken in Korea between 2013 and 2018. A broth microdilution antimicrobial susceptibility testing was performed for S. aureus. Molecular typing by multilocus sequence typing (MLST), spa typing, and pulsed-field gel electrophoresis (PFGE), was performed on mecA-positive S. aureus strain. S. aureus was isolated at a rate of 18.2% (777/4,264), of which MRSA comprised 0.7% (29 strains). MLST analysis showed that 11 out of the 29 MRSA isolates were predominantly sequence type (ST) 398 (37.9%). In addition, ST72, ST692, ST188, ST9, and ST630 were identified in the MRSA isolates. The spa typing results were classified into 11 types and showed a high correlation with MLST. The antimicrobial resistance assays revealed that MRSA showed 100% resistance to cefoxitin and penicillin. In addition, resistance to tetracycline (62.1%), clindamycin (55.2%), and erythromycin (55.2%) was relatively high; 27 of the 29 MRSA isolates exhibited multidrug resistance. PFGE analysis of the 18 strains excluding the 11 ST398 strains exhibited a maximum of 100% homology and a minimum of 64.0% homology. Among these, three pairs of isolates showed 100% homology in PFGE; these results were consistent with the MLST and spa typing results. Identification of MRSA at the final consumption stage has potential risks, suggesting that continuous monitoring of retail meat products is required.

LC-MS/MS Analysis of Surface Layer Proteins as a Useful Method for the Identification of Lactobacilli from the Lactobacillus acidophilus Group

  • Podlesny, Marcin;Jarocki, Piotr;Komon, Elwira;Glibowska, Agnieszka;Targonski, Zdzislaw
    • Journal of Microbiology and Biotechnology
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    • 제21권4호
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    • pp.421-429
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    • 2011
  • For precise identification of a Lactobacillus K1 isolate, LC-MS/MS analysis of the putative surface layer protein was performed. The results obtained from LTQ-FT-ICR mass spectrometry confirmed that the analyzed protein spot is the surface layer protein originating from Lb. helveticus species. Moreover, the identified protein has the highest similarity with the surface layer protein from Lb. helveticus R0052. To evaluate the proteomic study, multilocus sequence analysis of selected housekeeping gene sequences was performed. Combination of 16S rRNA sequencing with partial sequences for the genes encoding the RNA polymerase alpha subunit (rpoA), phenylalanyl-tRNA synthase alpha subunit (pheS), translational elongation factor Tu (tuf), and Hsp60 chaperonins (groEL) also allowed to classify the analyzed isolate as Lb. helveticus. Further classification at the strain level was achieved by sequencing of the slp gene. This gene showed 99.8% identity with the corresponding slp gene of Lb. helveticus R0052, which is in good agreement with data obtained by nano-HPLC coupled to an LTQ-FT-ICR mass spectrometer. Finally, LC-MS/MS analysis of surface layer proteins extracted from three other Lactobacillus strains proved that the proposed method is the appropriate molecular tool for the identification of S-layer-possessing lactobacilli at the species and even strain levels.

Ciprofloxacin 내성 대장균에서 Sequence Type과 Fluoroquinolone 내성의 분석 (Analysis of Sequence Type and Fluoroquinolone Resistance in Ciprofloxacin-Resistant Escherichia coli)

  • 조혜현
    • 대한임상검사과학회지
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    • 제53권3호
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    • pp.217-224
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    • 2021
  • 전 세계적으로 fluoroquinolone (FQ) 내성 그람음성균이 출현하고 있는 가운데, 최근 우리나라에서 FQ 내성 E. coli의 증가 추세는 심각한 우려를 낳고 있다. 이에 본 연구에서는 2018년 6월부터 12월까지 대전지역의 3차 병원에서 분리된 ciprofloxacin 내성 E. coli 56균주를 대상으로, 역학관계와 FQ 내성 결정인자의 양상을 조사하였다. 역학관계를 확인하기 위해 multilocus sequence typing (MLST)을 실시하였다. PCR과 염기서열 분석은 gyrA, gyrB, parC, parE 유전자의 QRDR에서 염색체상의 돌연변이와 aac(6)-Ib-cr, qepA, qnrA, qnrB, qnrC, qnrD 및 qnrS와 같은 PMQR 유전자의 빈도를 확인하였다. MLST 분석 결과, 12개의 ST를 확인하였으며, 이 중 가장 우세한 ST는 ST131 (31/56, 55.4%)이었고, 순차적으로 ST1193 (13/56, 23.2%), ST405 (3/56, 5.4%)의 결과를 보였다. ciprofloxacin 내성 E. coli 56균주 중 gyrA 유전자에서 83번째 아미노산인 serine (S)이 leucine (L)으로, 87번째 아미노산인 aspartic acid (D)가 asparagine (N)으로 치환되고, parC 유전자에서 80번째 아미노산인 serine (S)이 isoleucine (I)으로, 84번째 아미노산인 glutamic acid (E)가 valine (V)으로 치환된 결과(29/56, 51.8%)가 가장 빈번하게 확인되었고, aac(6)-Ib-cr (19/56, 33.9%)은 가장 흔한 PMQR 유전자로 확인되었다. 이러한 FQ 내성 결정인자의 결과는 다른 클론과 비교하여 ST131에서 더 빈번하게 확인되었다. ciprofloxacin 내성 E. coli 균주에 대한 역학적 특성의 지속적인 모니터링과 FQ 내성 결정인자에 대한 추가 연구가 필요할 것으로 사료된다.

Molecular Epidemiology of Clinical Cryptococcus neoformans Isolates in Seoul, Korea

  • Park, So Hae;Kim, Mina;Joo, Sei Ick;Hwang, Soo Myung
    • Mycobiology
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    • 제42권1호
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    • pp.73-78
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    • 2014
  • Cryptococcal infection is primarily caused by two species, Cryptococcus neoformans and C. gattii. Between the two species, C. neoformans var. grubii is the major causative agent of cryptococcosis in Asia. We investigated the molecular characteristics of 46 isolates of C. neoformans from patients with cryptococcosis between 2008 and 2012 in Seoul, Korea. All the isolates were determined to be C. neoformans var. grubii (serotype A), mating type $MAT{\alpha}$, and molecular type VNI by PCR-restriction fragment length polymorphism of the URA5 gene. Multilocus sequencing type (MLST) analysis using the International Society of Human and Animal Mycoses (ISHAM) consensus MLST scheme identified two sequence types (ST). Out of the 46 strains, 44 (95.7%) were identified as ST5, and remaining 2 were identified as ST31. Our study revealed that the clinical strains of C. neoformans in Korea are genetically homogeneous with the VNI/ST5 genotypes, and new appearance of VNI/ST31 genotype may serve as an important indicator of global genetic analysis.

Comparative Analysis of the Korean Population of Magnaporthe oryzae by Multilocus Microsatellite Typing

  • Choi, Jaehyuk;Kim, Hyojung;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • 제29권4호
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    • pp.435-439
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    • 2013
  • Rice blast fungus, Magnaporthe oryzae, inflicts serious damage to global rice production. Due to high variability of this fungal pathogen, resistance of newly-released rice cultivars is easily broken down. To understand the population structure of M. oryzae, we analyzed the genetic diversity of the Korean population using multilocus microsatellite typing. Eleven microsatellite markers were applied to the population of 190 rice isolates which had been collected in Korea for two decades since the 1980's. Average values of gene diversity and allele frequency were 0.412 and 6.5, respectively. Comparative analysis of the digitized allele information revealed that the Korean population exhibited a similar level of allele diversity to the integrated diversity of the world populations, suggesting a particularly high diversity of the Korean population. Therefore, these microsatellite markers and the comprehensive collection of field isolates will be useful genetic resources to identify the genetic diversity of M. oryzae population.