scholarly journals A molecular characterization ofClostridium difficileisolates from humans, animals and their environments

1993 ◽  
Vol 111 (2) ◽  
pp. 257-264 ◽  
Author(s):  
G. O'Neill ◽  
J. E. Adams ◽  
R. A. Bowman ◽  
T. V. Riley
Keyword(s):  
Fragment Length Polymorphism ◽  
Hospital Environment ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Direct Transmission ◽  
Chromosomal Dna ◽  
Veterinary Clinic ◽  
Enhanced Chemiluminescence ◽  
Typing Methods ◽  
Rflp Typing

SummaryIt is generally accepted that most patients withClostridium difficile-associated diarrhoea acquire the organism from the environment. Recently we demonstrated that household pets may constitute a significant reservoir ofC. difficilethrough gastrointestinal carriage in up to 39% of cats and dogs. These findings suggested that direct transmission from household pets, or contamination of the environment by them, may be a factor in the pathogenesis ofC. difficile-associated diarrhoea. To investigate this possibility, we examined isolates ofC. difficilefrom humans, pets and the environment by restriction enzyme analysis (REA) and restriction fragment length polymorphism (RFLP) typing using enhanced chemiluminescence. Both REA and RFLP typing methods usedHindIII digests of chromosomal DNA. A total of 116 isolates ofC. difficilefrom pets (26), veterinary clinic environmental sites (33), humans (37) and hospital environmental sites (20) was examined. REA was far more discriminatory than RFLP typing and for all isolates there were 34 REA types versus 6 RFLP types. There was good correlation between the REA types found in isolates from pets and from the veterinary clinic environment, and between isolates from humans and from those found in the hospital environment. There was, however, no correlation between REA type ofC. difficilefound in pets and isolates of human origin. We conclude that there may still be a risk of humans acquiringC. difficilefrom domestic pets as these findings may be the result of geographical variation.

Comparison of Restriction Enzyme Analysis, Arbitrarily Primed PCR, and Protein Profile Analysis Typing for Epidemiologic Investigation of an Ongoing Clostridium difficileOutbreak

1998 ◽  
Vol 36 (10) ◽  
pp. 2957-2963 ◽  
Author(s):  
Mary Ellen Rafferty ◽  
Aldona L. Baltch ◽  
Raymond P. Smith ◽  
Lawrence H. Bopp ◽  
Carol Rheal ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
General Hospital ◽  
Restriction Enzyme ◽  
Profile Analysis ◽  
Protein Profile ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Typing Methods ◽  
Arbitrarily Primed Pcr ◽  
Predominant Strain

During an outbreak of diarrhea in a general hospital in 1992, 166Clostridium difficile isolates from 102 patients were typed by restriction enzyme analysis (REA), arbitrarily primed PCR (AP-PCR), and protein profile analysis (PP) techniques. A total of 18 types and 5 subtypes were identified by REA, 32 types were identified by AP-PCR, and 9 types were identified by PP. Analysis of the data indicated the presence of a predominant strain among 76, 75, and 84% of the isolates by REA, AP-PCR, and PP, respectively. Subsequently, 45C. difficile isolates which had been collected in 1990 from 33 patients in the same hospital following a significant increase in the number of cases of diarrhea caused by C. difficile were studied by REA, AP-PCR, and PP typing techniques. Thirteen types and one subtype were identified by REA, 12 types were identified by AP-PCR, and 5 types were identified by PP. As with the isolates from 1992, a dominant strain was identified. This strain was represented by 53, 64, and 70% of the total number of isolates when the strains were typed by REA, AP-PCR, and PP, respectively. Every isolate (210 of 211) from both 1990 and 1992 that was available for typing was typeable by all three methods. Furthermore, the same dominant strain was identified in both 1990 and 1992 by each method. This study demonstrates that each of the three typing methods can be useful in epidemiologic investigations of C. difficileoutbreaks and that one strain can be dominant in an institution over a number of years.

scholarly journals Restriction enzyme analysis and ribotyping distinguish Bordetella avium and Bordetella hinzii isolates

2000 ◽  
Vol 124 (1) ◽  
pp. 83-90 ◽  
Author(s):  
R. E. SACCO ◽  
K. B. REGISTER ◽  
G. E. NORDHOLM
Keyword(s):  
Restriction Endonuclease ◽  
Restriction Enzyme ◽  
Restriction Endonucleases ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Bacterial Isolates ◽  
Chromosomal Dna ◽  
Bordetella Avium ◽  
Typing System

Fifty-seven bacterial isolates previously identified as Bordetella avium or B. hinzii were characterized by restriction enzyme analysis (REA) and/or ribotyping. Twenty restriction endonucleases were evaluated for REA. Digestion of chromosomal DNA from the 42 B. avium and 15 B. hinzii isolates with Hinf I produced 8 and 7 distinct fingerprint profiles, respectively. Digestion with DdeI further discriminated these Bordetella species and produced 12 fingerprint profiles for B. avium and 4 profiles of B. hinzii. In addition, B. avium isolates were clearly distinguishable from B. hinzii isolates by ribotyping with the restriction endonuclease PvuII. The ribotype patterns of these two species of Bordetella were unique when compared to previously reported ribotype patterns for B. bronchiseptica isolates. Since it was possible to discern differences among isolates within each Bordetella species by REA analysis, we suggest that REA could be used in developing a typing system based on the fingerprint profiles generated.

scholarly journals Repeated Recovery ofStaphylococcus saprophyticusFrom the Urogenital Tracts of Women: Persistence Vs. Recurrence

1995 ◽  
Vol 2 (5) ◽  
pp. 218-222 ◽  
Author(s):  
M. E. Rupp ◽  
J. Han ◽  
R. V. Goering
Keyword(s):  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Restriction Fragment Length Polymorphism ◽  
Tract Infection ◽  
Gel Electrophoresis ◽  
Fragment Length Polymorphism ◽  
Chromosomal Dna ◽  
Dna Restriction ◽  
Typing Methods ◽  
Restriction Fragment Length

Objective:The purpose of this study was to determine whether colonization was persistent or recurrent in a small group of women who had repeated recovery ofStaphylococcus saprophyticusfrom their urogenital tracts.Methods:Paired isolates ofS. saprophyticusfrom each of the study subjects were genotypically typed by plasmid fingerprinting and comparison of chromosomal-DNA restriction fragment-length polymorphism patterns by field-inversion gel electrophoresis (FIGE) and contour-clamped homogenous electric-field (CHEF) electrophoresis.Results:All isolates ofS. saprophyticusfrom the study subjects were classified as genetically unique by each of the typing methods.Conclusions:The subjects experienced recurrent colonization with different isolates ofS. saprophyticus.These findings may have broader implications regarding the pathogenesis and recurrence ofS. saprophyticusurinary-tract infection.

scholarly journals Use of Multiplex PCR and PCR Restriction Enzyme Analysis for Detection and Exploration of the Variability in the Free-Living Amoeba Naegleria in the Environment

2002 ◽  
Vol 68 (4) ◽  
pp. 2061-2065 ◽  
Author(s):  
Michel Pélandakis ◽  
Pierre Pernin
Keyword(s):  
Multiplex Pcr ◽  
Fragment Length Polymorphism ◽  
Internal Transcribed Spacers ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Polymorphism Analysis ◽  
Free Living ◽  
Pcr Product ◽  
Free Living Amoeba ◽  
Restriction Fragment Length

ABSTRACT A multiplex PCR was developed to simultaneously detect Naegleria fowleri and other Naegleria species in the environment. Multiplex PCR was also capable of identifying N. fowleri isolates with internal transcribed spacers of different sizes. In addition, restriction fragment length polymorphism analysis of the PCR product distinguished the main thermophilic Naegleria species from the sampling sites.

scholarly journals High-Resolution Genotyping ofCampylobacter Strains Isolated from Poultry and Humans with Amplified Fragment Length Polymorphism Fingerprinting

1999 ◽  
Vol 65 (6) ◽  
pp. 2369-2375 ◽  
Author(s):  
Birgitta Duim ◽  
Trudy M. Wassenaar ◽  
Alan Rigter ◽  
Jaap Wagenaar
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Amplified Fragment Length Polymorphism ◽  
Pcr Primers ◽  
Epidemiological Studies ◽  
Aflp Analysis ◽  
Campylobacter Coli ◽  
Chromosomal Dna ◽  
Typing Methods

ABSTRACT For epidemiological studies of Campylobacterinfections, molecular typing methods that can differentiate campylobacters at the strain level are needed. In this study we used a recently developed genotyping method, amplified fragment length polymorphism (AFLP), which is based on selective amplification of restriction fragments of chromosomal DNA, for genetic typing ofCampylobacter jejuni and Campylobacter colistrains derived from humans and poultry. We developed an automated AFLP fingerprinting method in which restriction endonucleasesHindIII and HhaI were used in combination with one set of selective PCR primers. This method resulted in evenly distributed band patterns for amplified fragments ranging from 50 to 500 bp long. The discriminatory power of AFLP was assessed with aC. jejuni strain, an isogenic flagellin mutant, and distinct C. jejuni strains having known pulsed-field gel electrophoresis and fla PCR-restriction fragment length polymorphism genotypes. Unrelated C. jejuni strains produced heterogeneous patterns, whereas genetically related strains produced similar AFLP patterns. Twenty-five Campylobacterstrains obtained from poultry farms in The Netherlands grouped in threeC. jejuni clusters that were separate from a C. coli cluster. The band patterns of 10 C. jejunistrains isolated from humans were heterogeneous, and most of these strains grouped with poultry strains. Our results show that AFLP analysis can distinguish genetically unrelated strains from genetically related strains of Campylobacter species. However, desirable genetically related strains can be differentiated by using other genotyping methods. We concluded that automated AFLP analysis is an attractive tool which can be used as a primary method for subtyping large numbers of Campylobacter strains and is extremely useful for epidemiological investigations.

scholarly journals Nosocomial Candida glabrata Colonization: an Epidemiologic Study

1998 ◽  
Vol 36 (2) ◽  
pp. 421-426 ◽  
Author(s):  
Jose A. Vazquez ◽  
Louise M. Dembry ◽  
Veronica Sanchez ◽  
Mary A. Vazquez ◽  
Jack D. Sobel ◽  
...  
Keyword(s):  
Candida Glabrata ◽  
Epidemiologic Study ◽  
Significant Risk ◽  
Marrow Transplant ◽  
University Hospital ◽  
Hospital Environment ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Indirect Contact ◽  
Environmental Surfaces

Candida glabrata has emerged as an important nosocomial pathogen, yet little is known about its epidemiology. We prospectively followed 98 patients admitted to a medical intensive care unit and the bone marrow transplant unit of a university hospital. Samples from environmental surfaces and the hands of hospital personnel were also cultured. Patients with newly acquired C. glabrata strains were compared to controls who were culture negative for C. glabrata. C. glabrata was recovered from multiple sites from 24 patients and three environmental surfaces. Sixteen patients (17%) acquired C. glabrata after admission to the study units. Significant risk factors for the nosocomial acquisition ofC. glabrata were prolonged duration of hospitalization in the unit and prior antimicrobial use. Strain delineation by restriction enzyme analysis revealed 28 different strains of C. glabrata; three strain types were common to nine patients. The environmental isolates were of the same strain type and common to five patients (four patients with newly acquired strains). These results suggest the possibility of exogenous nosocomial acquisition of C. glabrata, including the possible acquisition from the hospital environment. Transmission may be by indirect contact since identical strains of C. glabrata were recovered from patients who were geographically and temporally associated.

scholarly journals PCR-Restriction Enzyme Analysis of a Bone Marrow Isolate from a Human Immunodeficiency Virus-Positive Patient Discloses Polyclonal Infection with Two Mycobacterium avium Strains

2000 ◽  
Vol 38 (12) ◽  
pp. 4643-4645 ◽  
Author(s):  
R. S. Oliveira ◽  
M. P. Sircili ◽  
S. Y. M. Ueki ◽  
M. A. S. Telles ◽  
B. Schnabel ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mycobacterium Avium ◽  
Restriction Enzyme ◽  
Fragment Length Polymorphism ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Insertion Element ◽  
Immunodeficiency Virus ◽  
Digestion Pattern ◽  
Polyclonal Infection

Polyclonal infection by Mycobacterium avium was detected by hsp65 PCR-restriction enzyme analysis (PRA) in a bone marrow isolate from an AIDS patient. Two M. aviumstrains, differing in colony morphology, PRA HaeIII digestion pattern, insertion element (IS) 1245amplification, and restriction fragment length polymorphism fingerprints with IS1245 and IS1311 probes, were isolated.

scholarly journals Comparison of multilocus enzyme electrophoresis (MEE), ribotyping, restriction enzyme analysis (REA) and phage typing for typing ofListeria monocytogenes

1993 ◽  
Vol 111 (1) ◽  
pp. 71-79 ◽  
Author(s):  
B. Nørrung ◽  
P. Gerner-Smith
Keyword(s):  
Listeria Monocytogenes ◽  
Molecular Typing ◽  
Phage Typing ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Enzyme Electrophoresis ◽  
Typing Method ◽  
Multilocus Enzyme Electrophoresis ◽  
Serotype 1 ◽  
Typing Methods

SummaryThe discriminatory power of four methods for typing ofListeria monocytogeneswas compared. The four methods were multilocus enzyme electrophoresis (MEE), ribotyping, restriction enzyme analysis (REA), and a newly developed Danish phage typing system. Ninety-nine human clinical, food and slaughterhouse isolates ofListeria monocytogeneswere typed by each method. The most discriminatory single typing method was phage typing with an overall discriminatory index (DI) of 0·88 followed by REA, MEE and ribotyping with DI-values at 0·87, 0·83 and 0·79 respectively. Considering strains from each of the two predominant O-serotypes alone, serotype 1 was best discriminated by the molecular typing methods, in particular REA, which showed a DI of 0·92. The serotype 4 strains were best discriminated by phage typing (DI = 0·78). If two or more typing methods were combined, the combination of REA and MEE were found to be the most discriminatory combination. The DI values were 0·96, 0·74 and 0·90 for serotype 1, 4, and both combined, respectively. Phage typing is a rapid and inexpensive typing method but not as reproducible as the molecular typing methods. It is the most suitable method for mass screening. In situations where results are required to be highly reliable, i.e. when studying the relationships between only a few strains, a single or a combination of molecular typing methods should be used, preferable MEE and REA.

scholarly journals Relapse versus reinfection withClostridium difficile

1991 ◽  
Vol 107 (3) ◽  
pp. 627-635 ◽  
Author(s):  
G. L. O'Neill ◽  
M. H. Beaman ◽  
T. V. Riley
Keyword(s):  
Gastrointestinal Tract ◽  
Clostridium Difficile ◽  
Primary Culture ◽  
Restriction Enzyme ◽  
Rare Event ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Chromosomal Dna ◽  
Culture Plate ◽  
Multiple Strains

SUMMARYRelapse ofClostridium, difficile-associated diarrhoea occurs in 15–20% of patients; however, whether relapse is due to an endogenous source of the organism or reinfection from the environment remains unclear. Restriction enzyme analysis (REA) of chromosomal DNA was used to type multiple isolates from ten patients who had experienced apparent relapses. More than half the relapses were due to infection with a new strain ofC. difficile. The remaining patients were infected with the same strain, but whether this strain was acquired from the environment or from endogenous sources could not be determined. Relapses with a different strain ofC. difficilecould occur if an individual harboured more than one strain in their gastrointestinal tract. To investigate this possibility ten other patients were assessed for carriage of multiple strains. Ten colonies from a primary culture plate from each patient were typed by REA and tested for their ability to produce cytotoxin. All isolates from the same patient were identical by both methods, indicating that multiple carriage of strains may be a rare event.

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