• Title/Summary/Keyword: Ribotyping

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Enhanced Discrimination of Listeria spp. Using RAPD Fingerprinting Complemented by Ribotyping-PCR (리스테리아균의 특성분석을 위한 Molecular Typing 방법의 상호보완)

  • 임형근;홍종해;박경진;최원상
    • Journal of Life Science
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    • v.13 no.5
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    • pp.699-704
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    • 2003
  • The results typed by random amplification of polymorphic DNA (RAPD) were compared with those obtained by Enterobacterial repititive intergenic consensus (ERIC) fingerprinting and ribotyping-PCR. The discriminatory power of RAPD typing was the best among the methods tested. RAPD typing with two different primers for 13 Listeria spp. reference strains produced 11 patterns each. In contrast, ERIC fingerprinting produced 9 patterns and ribotyping-PCR produced 7 patterns each. Composite of two separate RAPD (Lis 11 and primer 6) results or RAPD (Lis11)/ ribotyping-PCR differentiated all 13 Listeria spp. reference strains. Therefore, composite of 2 separate RAPD (Lis11 and primer 6) or composite of RAPD (Lis11)/ribotyping-PCR is expected the most promising approach for typing field isolated Listeria spp. strains.

Molecular Epidemiology of Listeria monocytogenes by Ribotyping

  • Yang, Byoung-Seon
    • Biomedical Science Letters
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    • v.8 no.2
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    • pp.77-82
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    • 2002
  • Ten Listeria monocytogenes were isolated from clinical specimens and mussels, and their physio-biochemical characters were compared with the type strains. Ribotyping was used as a taxonomic tool to determine molecular epidemiological marker. Chromosomal DNA was cleaved with restriction enzymes HindIII and EcoRI. The fragment were subjected to Southern blot hybridization with 165 rDNA from B. subtilis by PCR. EcoRI patterns of Listeria strains showed 6 to 8 bands ranging from 0.75 kb to 11 kb band and they were classified into 6 groups. In comparison, HindIII patterns revealed that 5 to 7 bands ranging from 2.75 kb to 7.75 kb band and they classified into 5 groups. The various patterns of Listeria strains were observed within genus, species and isolated sources. 165 rRNA gene restriction patterns (ribotyping) are useful in epidemiological and taxonomic study.

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Comparison of Biotyping, Serotyping and Molecular Typing of Yersinia enterocolitica Isolated from Spring water in Seoul (서울시내 약수에서 분리한 Yersinia enterocolitica의 생물형, 혈청형 및 분자학적 형별비교)

  • 이영기;최성민;오수경;신재영
    • Journal of environmental and Sanitary engineering
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    • v.14 no.4
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    • pp.99-109
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    • 1999
  • Enteropathogenic Yersina enterocolitica is an important cause of human and animal disease. Phenotypic and genotypic characteristics currently used to identify Yersinia enterocolitica are not necessarily sufficient to differentiate pathogenic from non-pathogenic strains or to analyze the epidemiology of yersiniae at a molecular level. To improve the characterization of Yersinia enterocolitica, A total of 65 isolates of Yersinia enterocolitica were examined with bioserotyping, antibiotic susceptibilities, PFGE, PCR-ribotyping. Genomic DNA pattern generated by PFGE are highly specific for different strains of an organism and have significant value in epidemiologic investigations. The PFGE analysis of Not I-digested chromosomal DNA of Y. enterocolitica were performed with a CHEF Mapper(Bio-Rad, USA). Not I generated 19 restriction endonuclease digestion profiles(REDP). PCR-ribotyping, performed with primers complementry to conserved regions of 16S and 23S rRNA gene, generated 13 ribotypes. PCR-ribotyping can be considered a good technich for subtyping strains of Y.enterocolitica.

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Ribotyping of Porphyromonas Gingivalis Isolated from Rapidly Progressive Periodontitis Patients (급속진행성치주염 환자로부터 배양분리한 Porphyromonas gingivalis 균주의 ribotyping)

  • Kim, Jin-Hong;Choi, Bong-Kyu;Choi, Seong-Ho;Cho, Kyoo-Sung;Chai, Jung-Kiu;Kim, Chong-Kwan
    • Journal of Periodontal and Implant Science
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    • v.29 no.4
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    • pp.963-979
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    • 1999
  • This study examined ribotypes of 36 P. gingivalis strains isolated from 10 rapidly progressive periodontitis patients in Korean and revealed the presence of genetic heterogeneity among the patients. Ribotyping was performed by using a oligonucleotide probes based on 16S rRNA after whole genomic DNA had been digested with the restriction endonuclease enzyme Kpn I and Pst I. In addition, the antigenic heterogeneity of fimbrillin and protease activity was analysed to observe the virulency of P. gingivalis. The results were as follows. 1. Using KpnI, 6 ribotypes were detected, whereas 7 ribotypes were identified by using PstI. When combined two enzymes, a total of 8 ribotypes was subgrouped. 2. Ribotype I/e was the most common and detected in 4 among 10 patients. 3. The fimbrillin expressed from P. gingivalis isolates had the molecular size of 41kDa, 43kDa, 49kDa. It was observed that the size of fimbrillin with the same ribotypes could be identical. 4. All the P. gingivalis strains showed strong proteolytic activity and had the molecular size more than 120kDa. In summary, total 8 ribotypes were observed for isolates from rapidly progressive periodontitis patients. Forty percent of the patients harbored isolates exhibiting the same ribotype I/e, and it was observed that more than one ribotype can coexist in an individual patient.

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Screening of Lactobacilli Derived from Fermented Foods and Partial Characterization of Lactobacillus casei OSY-LB6A for Its Antibacterial Activity against Foodborne Pathogens

  • Chung, Hyun-Jung;Yousef, Ahmed E.
    • Preventive Nutrition and Food Science
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    • v.14 no.2
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    • pp.162-167
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    • 2009
  • Various fermented foods were screened in search of food-grade bacteria that produce bacteriocins active against Gram-negative pathogens. An isolate from a mold-ripened cheese presented antibacterial activity against Gram-positive and Gram-negative bacteria. The most active isolate was identified as Lactobacillus casei by a biochemical method, ribotyping, and membrane lipid analysis, and was designated as OSY-LB6A. The cell extracts of the isolate showed inhibition against Escherichia coli p220, E. coli O157, Salmonella enerica serovar Enteritidis, Salmonella Typhimurium, and Listeria monocytogenes. The antibacterial nature of the cell extract from the isolate was confirmed by eliminating the inhibitory effects of acid, hydrogen peroxide, and lytic bacteriophages. The culture supernatant and cell extract retained antibacterial activity after heating at $60{\sim}100^{\circ}C$ for $10{\sim}20$ min. The activity of the cell extract from Lb. casei was eliminated by pronase and lipase. Finally, the cell extract showed a bactericidal mode of action against E. coli in phosphate buffer solution, but it was bacteriostatic in broth medium and food extracts.

Riboprint and Virulence Gene Patterns for Bacillus cereus and Related Species

  • Kim, Young-Rok;Batt, Carl A.
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1146-1155
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    • 2008
  • A total of 72 Bacillus cereus strains and 5 Bacillus thuringiensis strains were analyzed for their EcoRI ribogroup by ribotyping and for the presence or absence of seven virulence-associated genes. From these 77 strains, 42 distinctive ribogroup were identified using EcoRI, but the two species could not be discriminated by their EcoRI ribogroup. The 77 strains were also examined by PCR for the presence of seven virulence-associated genes, cerAB, pi-plc, entFM, bceT, hblA, hblC, and hblD. All five Bacillus thuringiensis strains were positive for these genes. Although differences in the patterns of virulence genes were observed among the different B. cereus strains, within any given ribogroup the patterns of the seven virulence genes was the same. Pulsed-field gel electrophoresis (PFGE) analysis in combination with available chromosomal maps for a selected group of B. cereus strains revealed significant differences in their chromosome size and the placement of virulence genes. Evidence for significant rearrangements within the B. cereus chromosome suggests the mechanism through which the pattern of virulence-associated genes varies. The results suggest linkage between ribogroups and virulence gene patterns as well as no apparent containment of the latter within any particular species boundary.

Characterization of Silver Nanoparticles Synthesized by Using Marine Isolate Streptomyces albidoflavus

  • Prakasham, Reddy Shetty;Kumar, Buddana Sudheer;Kumar, Yannam Sudheer;Shankar, Guntuku Girija
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.614-621
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    • 2012
  • Silver nanoparticles production by the green chemistry approach was investigated using an isolated marine actinomycetes strain. The isolated strain was identified as Streptomyces albidoflavus based on chemotaxonomic and ribotyping properties. The strain revealed production of silver nanoparticles both extracellular and intracellularly. Surface Plasmon Resonance analysis with the function of time revealed that particle synthesis by this strain is reaction time dependent. The produced particles were spherical shaped and monodispersive in nature and showed a single surface plasmon resonance peak at 410 nm. Size distribution histograms indicated production of 10-40-nm-size nanoparticles with a mean size of 14.5 nm. FT-IR spectra of nanopartilces showed N-H, C-H, and C-N stretching vibrations, denoting the presence of amino acid/peptide compounds on the surface of silver nanoparticles produced by S. albidoflavus. Synthesized nanoparticles revealed a mean negative zeta potential and electrophoretic mobility of -8.5 mV and -0.000066 $cm^2/Vs$, respectively. The nanoparticles produced were proteinaceous compounds as capping agents with -8.5 mV zeta potential and revealed antimicrobial activity against both Gram-negative and -positive bacterial strains. Owing to their small size, these particles have greater impact on industrial application spectra.