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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The typing of Campylobacter is relatively poorly developed compared to that of the Enterobacteriaceae, and new molecular methods may provide useful approaches. The polymerase chain reaction was used to amplify randomly primed genomic DNA from Campylobacter isolates with an optimised randomly amplified polymorphic DNA protocol. Groups of isolates were analysed from chicken house environmental sources, chicken joints from retail sources, patients suffering from clinical disease and laboratory culture collections. Amplicons were separated by agarose gel electrophoresis, stained with ethidium bromide, and banding patterns captured in a digital form for computer analysis with GelCompar software. The method gave 100% typability and reproducibility for the isolates investigated and proved a useful technique for the epidemiological analysis of Campylobacter. Computer-based analysis of the randomly amplified polymorphic DNA generated profiles allowed relationships between isolates to be studied at the molecular level resulting in some indication of molecular correlates of the origins of isolates.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Random amplification of polymorphic DNA (RAPD) was evaluated as a genotypic method for typing clinical strains of Propionibacterium acnes. RAPD can suffer from problems of reproducibility if parameters are not standardised. In this study the reaction conditions were optimised by adjusting template DNA concentration and buffer constituents. All isolates were typeable using the optimised RAPD protocol which was found to be highly discriminatory (Simpson's diversity index, 0.98) and reproducible. Typing of P. acnes by optimised RAPD is an invaluable tool for the epidemiological investigation of P. acnes for which no other widely accepted method currently exists.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 197 (2001), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Whereas several important virulence factors in Escherichia coli O157 have been identified, studies suggest they are not always essential and are probably insufficient to account for the severe clinical manifestation of E. coli O157 infection. Identification of putative virulence determinants is crucial to the understanding of bacterial pathogenesis and genomic comparison analysis may aid the characterisation of unidentified virulence attributes. In this study, representational difference analysis (RDA) was used for genomic comparison of E. coli O157 with the proposed ancestral strain, E. coli O55. Unique E. coli O157 gene sequences were isolated and one, termed RDA-1, taken forward for further analysis. Southern blotting with labelled RDA-1 as a probe showed it to be present in 77% of E. coli O157 isolates and absent in all non-E. coli O157 screened. Sequence flanking RDA-1 was obtained from a genomic clone identified by hybridisation, and contained an open reading frame predicted to encode a novel iron-regulated outer membrane protein.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The number, diversity and restriction enzyme fragmentation patterns of plasmids harboured by 44 multidrug-resistant hospital-acquired methicillin-resistant Staphylococcus aureus (MR-HA-MRSA) isolates, two multidrug-resistant community-acquired MRSA (MR-CA-MRSA), 50 hospital-acquired MRSA (HA-MRSA) isolates (from the University Hospital Birmingham, NHS Trust, UK) and 34 community-acquired MRSA (CA-MRSA) isolates (from general practitioners in Birmingham, UK) were compared. In addition, pulsed-field gel electrophoresis (PFGE) type following SmaI chromosomal digest and SCCmec element type assignment were ascertained for each isolate. All MR-HA-MRSA and MR-CA-MRSA isolates possessed the type II SCCmec, harboured no plasmid DNA and belonged to one of five PFGE types. Forty-three out of 50 HA-MRSA isolates and all 34 CA-MRSA isolates possessed the type IV SCCmec and all but 10 of the type IV HA-MRSA isolates and nine CA-MRSA isolates carried one or two plasmids. The 19 non-multidrug-resistant isolates (NMR) that did not harbour plasmids were only resistant to methicillin whereas all the NMR isolates harbouring at least one plasmid were resistant to at least one additional antibiotic. We conclude that although plasmid carriage plays an important role in antibiotic resistance, especially in NMR-HA-MRSA and CA-MRSA, the multidrug resistance phenotype from HA-MRSA is not associated with increased plasmid carriage and indeed is characterised by an absence of plasmid DNA.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    World journal of microbiology and biotechnology 16 (2000), S. 741-748 
    ISSN: 1573-0972
    Keywords: AFLP ; epidemiological typing ; Escherichia coli O157 ; molecular epidemiology ; PFGE ; RAPD ; sub-typing ; typing methods ; VTEC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The epidemiological investigation of Escherichia coli O157 is complicated by the lack of heterogeneity between strains responsible for the majority of cases of infection. As a consequence it is difficult to reliably cluster together an outbreak strain and differentiate it from other sporadically occurring isolates. The methods available for the sub-typing of E. coli O157 vary in their speed, technical complexity, cost and ability to discriminate reliably between strains, with many of the recently developed methods targeting the genome to provide differentiation. No single typing method is individually superior, and ideally a combination of techniques should be employed depending on the level of discrimination required and time or resources available. The aim of this review is to consider the relative merits of the available typing methodologies with particular emphasis on those which may find application in a diagnostic laboratory.
    Type of Medium: Electronic Resource
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