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B D Cookson

Publications and source records attributed to B D Cookson.

At least 55 records · Page 3Linked to original sources

Comparison of serotype, biotype and bacteriocin type with rDNA RFLP patterns for the type identification of Serratia marcescens.

Variations in rDNA gene loci in DNA digests of 209 clinical isolates of Serratia marcescens were determined with an Escherichia coli rRNA probe. Forty-one restriction fragment length polymorphism patterns (ribotypes) were identified, based on the size of 4-14 (mean 7.5) hybridization bands. The patterns differed by more than a single band in 98% of pair-wise comparisons. On a subset of 76 isolates, ribotyping proved to be marginally more discriminating than biotyping (discrimination index 0.92 v. 0.89) followed by serotyping (0.87) and bacteriocin typing (0.74). About one-third of isolates belonged to unique ribotypes and only two ribotypes exceeded 5% in frequency (23.0 and 6.4% respectively). A combination of serotype or biotype with ribotyping defined a similar number of strains, although none of the methods alone was sufficiently discriminatory to identify strains. We conclude that due to the accessibility of biotyping and the lack of commercially available antisera for S. marcescens, the biotype and ribotype together provide reliable markers of strain identity.

Bacterial Typing Techniques↗

Phenotypic and phylogenetic characterization of some Gemella-like organisms from human infections: description of Dolosigranulum pigrum gen. nov., sp. nov.

A phylogenetic analysis was performed on two previously uncharacterized Gram-positive, catalase-negative bacteria from clinical sources. 16S rRNA sequencing studies revealed the isolates represent a new line of descent within the lactic acid group of bacteria. On the basis of the phylogenetic findings and phenotypic distinctiveness of the organisms, it is proposed that they be classified in a new genus Dolosigranulum, as Dolosigranulum pigrum sp. nov. The type strain of Dolosigranulum pigrum is NCFB 2975.

Base Sequence↗

An epidemic methicillin-resistant strain of Staphylococcus aureus in Spain.

During 1990, a strain of methicillin-resistant Staphylococcus aureus became epidemic in Spain and spread in a manner analogous to that of EMRSA-1 in England. Isolates of this strain produced little protein A and were resistant to a number of antibiotics including ciprofloxacin. Beta-lactamase production was encoded by a c. 39 kb plasmid, which also conferred resistance to mercury, cadmium, ethidium bromide and propamidine isethionate. Investigation showed that two variants, separable by supplementary and Fisk phage typing, were circulating. The B variant appeared to spread more readily than the A variant. The opportunity was taken to compare the discriminatory power of traditional typing methods with molecular techniques. The discriminatory power of the molecular techniques used only reached the same level as the traditional methods when double enzyme digestion of total cellular DNA by EcoR I and Cla I was performed.

Bacterial Typing Techniques↗

Ribotyping of coagulase-negative staphylococci.

The discriminative capacity of ribotyping was initially assessed without knowledge of results obtained for the same isolates by use of more established typing methods. Forty-eight isolates of coagulase-negative staphylococci (CNS) from peritoneal fluids were studied. They were collected prospectively during 31 consecutive episodes of infection associated with peritoneal dialysis in 17 patients. DNA was digested by the restriction endonucleases EcoRI or HindIII and ribotyped by means of a biotinylated cDNA probe to 16S + 23S staphylococcal ribosomal RNA gene sequences. These methods in combination produced a total of 27 types which compared well with numbers of groups distinguished by other typing methods: limited biotype-antibiotic resistogram (ARB; 28), antibiotic resistogram alone (25), API-Staph (12), phage typing (9) and plasmid analysis (22). Ribotyping was highly reproducible and typed all isolates, including those that were not phage-typable (35) or did not contain plasmids (4). When used in a hierarchical manner with ARB, ribotyping results produced 13 additional types in comparison with the other three methods. When used hierarchically with all other typing systems, a further five types were found among isolates from two patients. However, some of the differences observed as a result of ribotyping could have been due to subtle changes produced by mutation, lysogenisation or gene transposition. Since the method requires additional time, expense and technical expertise, it is likely to be useful only when answers to specific epidemiological problems are required or as an initial screen before using other methods of genetic analysis.

Base Sequence↗

Screening for Corynebacterium diphtheriae.

A throat swab from a 9 year old girl with pharyngitis yielded a non-toxigenic strain of Corynebacterium diphtheriae var mitis and Streptococcus group G. C pseudodiphtheriticum was isolated from the throats of two of her four brothers. In each case the isolate was sent to the reference laboratory before full identification. The growth was found to be mixed for one brother; the other isolate being a toxin producing C diphtheriae var gravis. The child was asymptomatic and the case proves that all colonial types on the Hoyles plate should be identified.

Child↗

The epidemiology of ciprofloxacin resistance in coagulase-negative staphylococci in CAPD patients.

Ciprofloxacin was used as empirical therapy for peritonitis in patients receiving continuous ambulatory peritoneal dialysis (CAPD) for 26 months, providing an opportunity to study the epidemiology of ciprofloxacin resistance amongst coagulase-negative staphylococci (CNS). Swabs were collected from the CAPD patients, staff, and clinic environment before, during and after the time this antibiotic was prescribed. Clinical isolates were also studied, and records kept of patient hospital attendance. Ciprofloxacin-resistant staphylococci were typed by antibiogram, biotype, plasmid profile, SDS-PAGE, and immunoblotting. No resistant strains were detected before the use of ciprofloxacin. During its use 30% of patients became skin carriers, and resistant strains caused 8% of peritonitis episodes in 7% of patients (38% of carriers). Resistant strains were isolated from the environment, but never from attending members of staff. A total of 208 resistant isolates with MIC's between 8 and 128 mg/l was collected and identified as Staphylococcus epidermidis or S. haemolyticus. Sixteen strain types were distinguished. There was epidemiological evidence for selection of resistant strains derived from the host commensal flora and also for cross-colonization, and cross-infection. Carriage of resistant strains fell to 15% of patients, 6 months after the use of ciprofloxacin had ceased.

Air Microbiology↗

Inter-strain comparison by pyrolysis mass spectrometry in the investigation of Staphylococcus aureus nosocomial infection.

Pyrolysis mass spectrometry (PyMS) was used to examine isolates of Staphylococcus aureus from an outbreak of wound infections on a cardiothoracic surgical unit, some of which were thought to have been related to a point-source in the operating theatre. The PyMS results were compared with the results of phage typing. Both methods suggested that a single strain of S. aureus, of phage pattern 29/52/52A/79/80/81, was responsible for some of the wound infections, but PyMS also identified two patients with phage non-typable isolates. Phage typing indicated four staff members as possible carriers of the epidemic strain, but PyMS indicated only two. Epidemiological enquiry confirmed that one of the two members of staff identified by both methods was likely to have been the source of the theatre-based infection. PyMS is a rapid and relatively inexpensive technique for the investigation of nosocomial S. aureus infection and was more discriminatory than phage typing in this instance.

Carrier State↗

Chlorhexidine resistance in methicillin-resistant Staphylococcus aureus or just an elevated MIC? An in vitro and in vivo assessment.

Chlorhexidine (Hibiscrub; ICI) is generally accepted to be effective as an antiseptic hand wash for methicillin-susceptible Staphylococcus aureus (MSSA), but there is dispute whether the chlorhexidine MIC for methicillin-resistant S. aureus (MRSA) strains is higher than that for MSSA strains and, indeed, whether it is relevant. In addition, the link between resistance to chlorhexidine, gentamicin, and "nucleic acid-binding" compounds (NAB; which code, in particular, for propamidine isethionate and ethidium bromide) requires clarification. We performed chlorhexidine MIC and rate of kill tests on a number of MSSA and MRSA isolates. Two gentamicin-resistant MRSA isolates without NAB plasmids were more susceptible (0.25 and 0.5 microgram/ml) than four of eight MSSA that we tested (range, 0.25 to 2 microgram/ml). Chlorhexidine MICs were higher (4 to 8 micrograms/ml) for seven distinct MRSA isolates with plasmids conveying resistance to gentamicin and NAB (GNAB). Curing of the GNAB plasmid from MRSA strains resulted in a fall in the MIC (1 to 3.3 micrograms/ml), but no consistent fall in killing by chlorhexidine was observed. No effect on the chlorhexidine MIC or killing was observed when we cured strains of methicillin resistance. GNAB plasmid transfer resulted in a rise in the chlorhexidine MIC for the strains but not consistent fall in killing by chlorhexidine. Ethical approval was granted for 10 volunteers to each have a methicillin-susceptible, GNAB-resistant, derived transcipient and its GNAB-susceptible isogenic parent applied to separate sites in an in vivo skin test; no significant difference was seen in survival rates after the application of chlorhexidine. These results suggest that chlorhexidine appears to be as effective as a hand-washing agent for MRSA isolates with or with out NAB plasmids as it is for MSSA isolates.

Chlorhexidine↗