Comparative evaluation of an automated ribotyping instrument versus pulsed-field gel electrophoresis for epidemiological investigation of clinical isolates of bacteria

Hollis, Richard J.; Bruce, James L.; Fritschel, S. J.; Pfaller, Michael A.

doi:10.1016/s0732-8893(99)00033-4

Cited by 61 publications

(46 citation statements)

References 11 publications

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“…Each lane of sample data was normalized to a standard marker set and band intensity, and then compared with reference patterns. Similarity coefficients were calculated based on both position and relative band weight (17,22).…”

Section: Methodsmentioning

confidence: 99%

Stpahylococcus aureus biofilms on central venous haemodialysis catheters

Aoki¹,

Pizzolitto²,

Garcia³

et al. 2005

Braz. J. Microbiol.

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Biofilm bacterial infections are common in patients undergoing treatment with haemodialysis. This study involved 16 patients (7 males, 9 females; ages from 22 to 81 with an average age of 50) who had had a total of 25 temporary haemodialysis polyurethane catheter insertions into the subclavian vein (22 dual-lumen and 3 triple-lumen). The catheters remained in place from 3 to 91 days, on an average of 47 days. The reasons for catheter removal were: bad functioning (44%), suspicion of catheter-related infection (20%), availability of permanent access (16%), accidental removal (12%), signs and symptoms of infection at the site of catheter insertion (4%), and exogenous contamination (4%). Positive tip cultures were observed on seven of the catheters (28%), showing three positive blood cultures. The Staphylococcus aureus were identified in 12% of the blood cultures and isolated from one of the hubs, and biofilms were observed on all catheter tips. The S. aureus retrieved from both blood and catheters (tips and hubs) were resistant to penicillin and susceptible to azithromycin, ciprofloxacin, clindamycin, chloramphenicol, gentamicin, oxacillin, rifampin, sulfamethoxazole, tetracycline, and vancomycin. The S. aureus strains isolated from both blood and catheters (tips and hubs) were considered to be identical based on antibiotic susceptibility patterns and genetic similarity assessed using an automated ribotyping system.

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Section: Methodsmentioning

confidence: 99%

Stpahylococcus aureus biofilms on central venous haemodialysis catheters

Aoki¹,

Pizzolitto²,

Garcia³

et al. 2005

Braz. J. Microbiol.

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“…Herein, we describe the use of pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), ribotyping, serotyping, and virulence gene detection by PCR to study in detail the molecular epidemiology of V. parahaemolyticus strains isolated from Canadian patients (3,(9)(10)(11). A retrospective descriptive laboratory analysis of clinical V. parahaemolyticus isolates from across Canada (primarily British Columbia), which is available from the inventory of the National Microbiology Laboratory, Public Health Agency of Canada, was conducted.…”

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confidence: 99%

Phenotypic and Genotypic Characterization of Canadian Clinical Isolates of Vibrio parahaemolyticus Collected from 2000 to 2009

et al. 2014

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e Vibrio parahaemolyticus is the leading bacterial cause of food-borne illness due to the consumption of contaminated seafood. The aim of the present study was to determine the population of its subtypes and establish a better understanding of the various types of V. parahaemolyticus strains that are causing human illness in Canada. The subtypes for 100 human clinical isolates of V. parahaemolyticus collected between 2000 and 2009 were determined by performing serotyping, ribotyping, pulsed-field gel electrophoresis, and multilocus sequence typing. Within this panel of strains, there was a high level of diversity (between 22 and 53 subtypes per method), but the presence of predominant clones with congruent subtypes between the various methods was also observed. For example, all 32 isolates belonging to sequence type 36 (ST36) were from serogroup O4, while 31 of them were ribotype EcoVib235-287, and 24 of the 32 were SfiI pulsed-field gel electrophoresis (PFGE) pattern VPSF1.0001. With regard to the presence of known virulence genes, 74 of the 100 isolates were PCR positive for the presence of the thermostable direct hemolysin (tdh); and 59 of these 74 strains also contained the second virulence marker, the tdh-related hemolysin (trh). The detection of trh was more predominant (81%) among the clinical isolates, and only four (4%) of the clinical isolates tested negative for the presence of both tdh and trh. This database, comprising 100 clinical isolates of V. parahaemolyticus strains from Canada, forms a baseline understanding of subtype diversity for future source attribution and other epidemiologic studies.

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“…), as described previously (5,16). Briefly, isolates were inoculated into tubes containing lysis buffer, placed in a heating block at 80°C for 30 min, and then transferred to the RiboPrinter instrument.…”

mentioning

confidence: 99%

“…Genomic DNA in agarose was digested with the restriction enzyme PvuII, and the resulting fragments were separated by electrophoresis in 1% agarose on a CHEF-DRII apparatus (Bio-Rad, Richmond, Calif.) with the following conditions: 200 V for 23 h at switch times ramped from 5 to 40 s. Strains were considered different by PFGE if more than three bands were different (5,21). Strains with one to three different bands were considered subtypes.…”

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confidence: 99%

Modification of Dienes Mutual Inhibition Test for Epidemiological Characterization of Pseudomonas aeruginosa Isolates

2002

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Pseudomonas aeruginosa is an important cause of community-associated and nosocomial infections related to exposure to aqueous environments. Such infections often occur in the setting of a common-source outbreak, in which case epidemiological characterization of isolates may be necessary. In this preliminary study, a modification of the Dienes mutual inhibition test, ordinarily used to assess the relatedness of swarming Proteus mirabilis strains, was used to study 15 P. aeruginosa isolates, with the results compared to those obtained by ribotype analysis. Complete concordance was noted between the results of the Dienes test and those of ribotyping. These observations suggest that further studies are warranted to assess the utility of the modified Dienes test as a simple, inexpensive, and reliable means for epidemiological typing of P. aeruginosa.Pseudomonas aeruginosa is a ubiquitous gram-negative bacillus that is often found in moist environments (6). Community-associated P. aeruginosa infections may occur as a consequence of exposure to the organism in hot tubs, whirlpools, water slides, or swimming pools (8) or by use of contaminated sponges (2, 11). Keratitis due to P. aeruginosa may occur in individuals who store contact lenses in tap water and/or contaminated contact lens solutions (4). Intravenous drug users may develop endocarditis and osteomyelitis as a result of injection of contaminated injectables (9).P. aeruginosa is also of nosocomial significance. The organism has been isolated from a variety of solutions including soaps, ointments, irrigation and dialysis fluids, eyedrops, and disinfectants, as well as from fomites such as showerheads, respiratory therapy equipment, sinks, and baths (12). Populations at risk for significant morbidity and mortality include intubated patients in intensive care units (17), patients on chronic ambulatory peritoneal dialysis (1), and burn patients (13).Epidemiological investigations of P. aeruginosa infection have used molecular typing methods (10), as well as tools such as biotyping, phage typing, bacteriocin typing, and serotyping (7). The Dienes mutual inhibition test has been used as an epidemiological tool to characterize isolates of Proteus mirabilis (15,18,19,20). In this test, if two different P. mirabilis colonies growing on an agar plate are genetically dissimilar, a clear line of demarcation forms at the point of intersection of swarming growth; conversely, if two isolates are identical or highly similar, the swarming edges merge without a conspicuous line of demarcation (3,18,19). Colonies of P. aeruginosa usually have a spreading morphology on agar media (8) and have the capability of adhering to central venous catheters (CVCs), including those impregnated with minocycline and rifampin (E. L. Munson et al., unpublished data). In this preliminary report, we propose a modified Dienes test for the epidemiological characterization of P. aeruginosa isolates.Ten clinical isolates of P. aeruginosa, previously identified by the Vitek GNIϩ card (bioMérieux, Marcy l...

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Comparative evaluation of an automated ribotyping instrument versus pulsed-field gel electrophoresis for epidemiological investigation of clinical isolates of bacteria

Cited by 61 publications

References 11 publications

Stpahylococcus aureus biofilms on central venous haemodialysis catheters

Stpahylococcus aureus biofilms on central venous haemodialysis catheters

Phenotypic and Genotypic Characterization of Canadian Clinical Isolates of Vibrio parahaemolyticus Collected from 2000 to 2009

Modification of Dienes Mutual Inhibition Test for Epidemiological Characterization of Pseudomonas aeruginosa Isolates

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