Characterisation of methicillin-resistant Staphylococcus aureus by biotyping, immunoblotting and restriction enzyme fragmentation patterns

Coia, John; Thomson-Carter, Fiona M.; Baird, D. R.; Platt, D. J.

doi:10.1099/00222615-31-2-125

Cited by 33 publications

(17 citation statements)

References 4 publications

(4 reference statements)

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“…An association between nasal carriage of S. aureus and subsequent infections was also described in studies evaluating patients on continuous ambulatory peritoneal dialysis (CAPD) [23]. On the bases of typing methods Coia et al [7] found identical isolates from the site of infection and from carriage sites. These results showed that for patients on CAPD a S. aureus carriage is an important risk factor for development of S. aureus infections.…”

Section: Discussionmentioning

confidence: 99%

The significance of nasal carriage of Staphylococcus aureus as risk factor for human skin infections

Toshkova

2001

FEMS Microbiology Letters

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The present study was designed to investigate the significance and the relationship between nasal carriage of Staphylococcus aureus and staphylococcal skin infections. Thirty-one S. aureus strains, isolated from 12 patients with chronic and recurrent skin infections, one patient with septicemia and one patient with otitis externa were studied. The staphylococcal strains were isolated from the site of infection and from the anterior nares of each patient. The identity of both strains of each pair could be demonstrated by determination of phenotypic properties and by genotyping of the isolates. The phenotypic properties included hemolytic activities, antibiotic resistance data, and the production of enterotoxins. The identity was additionally confirmed by phage-typing, by determination of the size and the number of repeats of the X region of spa gene, by determination of gene polymorphisms of coa gene and by macrorestriction analysis of the chromosomal DNA of the isolates by pulsed-field gel electrophoresis. The present results showed an identity of the S. aureus obtained from anterior nares and from skin infection of each patient indicating the importance of nasal carriage of these bacteria for development of human skin infection. ß

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Section: Discussionmentioning

confidence: 99%

The significance of nasal carriage of Staphylococcus aureus as risk factor for human skin infections

Toshkova

2001

FEMS Microbiology Letters

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“…Staphylococci were identified based on Gram reaction (Gram-positive cocci in clumps), catalase, coagulase (tube method using rabbit plasma) and DNase tests. Biotyping was based on the hydrolysis of urea, Tween 80 and pigmentation on Isosensitest agar (Coia et al, 1990). Pigmentation was classified as yellow (Y), cream (C) or white (W).…”

Section: Methodsmentioning

confidence: 99%

Identification and molecular characterization of mannitol salt positive, coagulase-negative staphylococci from nasal samples of medical personnel and students

Shittu

Lin

Morrison

et al. 2006

Journal of Medical Microbiology

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The identification of mannitol salt positive, coagulase-negative staphylococci (CNS) is often disregarded when Staphylococcus aureus is screened in clinical samples using mannitol salt agar. However, the emergence of CNS as important human pathogens has indicated that reliable methods for the identification of clinically significant CNS are of great importance in understanding the epidemiology of infections caused by them. The identification and molecular characterization of mannitol salt positive CNS from nasal samples of medical personnel and students is reported here. A total of 84 mannitol salt positive staphylococcal isolates were obtained from 240 nasal samples, of which 15 were CNS. The API STAPH system classified the CNS isolates into six species, and one-third of the isolates were identified with confidence levels of <80 %. 16S-23S rRNA intergenic spacer length polymorphism analysis (ITS-PCR) identified only two species (Staphylococcus haemolyticus and Staphylococcus saprophyticus). This identification was confirmed by antibiotyping, species-specific PCR and PFGE. The results from this study indicate that ITS-PCR is a potentially useful and reliable tool, enabling hospital laboratories to obtain rapid, full and accurate identification of CNS at the species level. INTRODUCTIONThe identification of bacterial pathogens in human infection plays a key role in the management of patients in health care institutions. In many clinical laboratories, Staphylococcus aureus is usually isolated on nonspecific media (e.g. blood agar) and then presumptively identified before definitive overnight characterization (Kloos & Bannerman, 1995). In an attempt to achieve presumptive isolation in a single step, mannitol salt agar (MSA) was developed in 1945 for the selective isolation of pathogenic staphylococci in the clinical microbiology laboratory (Chapman, 1945;Blair et al., 1967). The growth and production of yellow colonies, due to the high salt content of the medium and fermentation of mannitol, is regarded as a presumptive tool in the identification of S. aureus. It is also described as a characteristic for the differentiation of coagulase-positive staphylococci from coagulase-negative staphylococci (CNS) (Duguid, 1989). However, there are reports that some CNS can also produce yellow colonies on MSA (Martinez et al., 1992;Merlino et al., 1996;Mir et al., 1998;Jayaratne & Rutherford, 1999;Simor et al., 2001;Zadik et al., 2001). Mannitol salt positive CNS impede and delay the isolation and subsequent identification of S. aureus in clinical samples on the primary plate. Nevertheless, they have attracted very little attention and in most cases are not identified to species level.As a group, the CNS are among the most frequently isolated bacteria in the clinical microbiology laboratory. They are becoming increasingly important as causative agents of hospital-acquired bacteraemia, with the increasing use of prosthetic devices and other invasive technologies in medical institutions (von Eiff et al., 2002 The aim of this stud...

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“…These ranges from a traditional method like phage typing (Williams and Rippon, 1952), biotyping (Coia et al, 1990), serotyping (Melo Cristino et al, 1986), to classical methods, which make use of molecular biological techniques (pulse field gel electrophoresis (PFGE) (Wielders et al, 2002), ribotyping (Wielders et al, 2002), restriction fragment analysis of plasmid) such as looking at differences in chromosome and extra-chromosomal element like plasmid. This study was aimed in assessing the value of the detection of rDNA polymorphisms in the genome of S. aureus as a typing system in studying the epidemiology of MRSA.…”

Section: Methicillin Resistant Staphylococcus Aureusmentioning

confidence: 99%

A comparative assessment of ribosomal DNA polymorphisms in methicillin resistant Staphylococcus aureus (MRSA) epidemiology

Alli¹,

Akinloye²,

Rowley³

et al. 2010

African Journal of Biomedical Research

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Chromosomal DNA from 19 strains of methicillin resistant Staphylococcus aureus (MRSA) which included 13 strains from different geographical locations in England and 6 strains from St George's Hospital (SGH), London, with MRSA NCTC 119040 and Oxford S. aureus were digested with the restriction endonucleases, HindIII and HaeIII. Southern blot hybridisation was carried out on the DNA digests transferred onto membrane using radioactive probe prepared from 16S and 23S rRNA from Escherichia coli. The pattern of bands which depended on restriction fragment length polymorphisms was used as a measure of minor genomic variation within the MRSA strains. Phage typing, biotyping, antibiogram, and plasmid profile analysis were also carried out on all the strains for comparison. Four and five different patterns were obtained for HindIII and HaeIII, respectively for representative isolates from different geographical locations in England for that year. Three different patterns were identified among the MRSA strains from SGH, London, using HaeIII. None of the epidemiological markers used in this study gave results that correlated with each other. Ribosomal DNA polymorphisms have proved to be a very useful technique for studying the epidemiology of MRSA in hospitals when used along with other epidemiological markers.

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Characterisation of methicillin-resistant Staphylococcus aureus by biotyping, immunoblotting and restriction enzyme fragmentation patterns

Cited by 33 publications

References 4 publications

The significance of nasal carriage of Staphylococcus aureus as risk factor for human skin infections

The significance of nasal carriage of Staphylococcus aureus as risk factor for human skin infections

Identification and molecular characterization of mannitol salt positive, coagulase-negative staphylococci from nasal samples of medical personnel and students

A comparative assessment of ribosomal DNA polymorphisms in methicillin resistant Staphylococcus aureus (MRSA) epidemiology

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