Hybridization of EcoRI- and HindIII-digested chromosomal DNAs from 41 isolates of Enterococcus faecalis with probes for rRNA genes was performed (ribotyping). The ability of ribotyping to distinguish strains at the subspecies level was compared with results previously determined by pulsed-field gel electrophoresis (PFGE). With EcoRI, seven ribopatterns (usually differing by only one band) were found, while PFGE had previously shown 25 clearly different patterns plus six related variants. Digestion with HindIII generated a few additional patterns but still failed to differentiate some strains that had very different PFGE patterns. Ribotyping with BscI has also been reported to be inadequate for subspecies strain differentiation (L. M. Hall, B. Duke, M. Guiney, and R. Williams, J. Clin. Microbiol. 30:915-919, 1992). Although ribotyping with other restriction endonucleases may perform better in distinguishing different strains, at present PFGE appears to be superior for strain differentiation. ...
TY - JOUR. T1 - Assessment of resolution and intercenter reproducibility of results of genotyping Staphylococcus aureus by pulsed-field gel electrophoresis of SmaI macrorestriction fragments. T2 - A multicenter study. AU - Van Belkum, Alex. AU - Van Leeuwen, Willem. AU - Kaufmann, Mary Elizabeth. AU - Cookson, Barry. AU - Forey, Françoise. AU - Etienne, Jerome. AU - Goering, Richard V.. AU - Tenover, Fred. AU - Steward, Christine. AU - OBrien, Frances. AU - Grubb, Warren. AU - Tassios, Panayotis. AU - Legakis, Nicholas. AU - Morvan, Anne. AU - El Solh, Névine. AU - De Ryck, Raf. AU - Struelens, Marc. AU - Salmenlinna, Saara. AU - Vuopio-Varkila, Jaana. AU - Kooistra, Mirjam. AU - Talens, Adriaan. AU - Witte, Wolfgang. AU - Verbrugh, Henri. PY - 1998/6. Y1 - 1998/6. N2 - Twenty well-characterized isolates of methicillin-resistant Staphylococcus aureus were used to study the optimal resolution and interlaboratory reproducibility of pulsed-field gel electrophoresis (PFGE) of DNA macrorestriction ...
Electrophoreseis is usually carried out within a matrix or gel made of agarose or polyacrylamide. These gels are chemically inert, so they will interfere little with the molecules. The sample is loaded in the gel;in wells for nucleic acids separation. Agarose is a polysaccharide extracted from seaweed. Agarose gels have a large range of separation depending on the concentration.The higher the concentration the smaller the pore size will be.Polyacrylamide is a cross-linked polymer of acrylamide.To avoid inhibition of polymerization by oxygen, they are poured between glass plates to mage gel slabs.Low concentration of polyacrylamide or less cross linking results in gels with large pores. Standard protein gels are typically composed of two layers, ahe top-most layer called the stacking gel and a lower layer called separating or resolving gel.The stacking layer contains a low percentage of acylamide and low pH , while the acrylamide concentration of the separating gel varies according to the samples ...
Pulsed-field gel electrophoresis (PFGE) was performed with a contour-clamped homogeneous electric field DRII apparatus from Bio-Rad Laboratories (Richmond, Calif.) as described previously (7). The chromosomal DNA was digested overnight with XbaI (GIBCO-BRL, Life Technologies, Gaithersburg, Md.). DNA was electrophoresed in 1.2% SeaKem GTG agarose (FMC) at 6 V/cm for 24 h; the pulse time was increased from 5 to 40 s. Because a single base mutation in the chromosomal DNA of an isolate is sufficient to introduce differences in three fragments in its restriction pattern, isolates with restriction patterns showing the same differences in one to three fragments were considered to belong to the same genotype (23). The PFGE patterns were also analyzed with the computer software Gelcompar for Windows version 3.1b (Applied Math, Kortrijk, Belgium). The PFGE patterns were compared by the algorithmic clustering method called the unweighted-pair group method using arithmetic averages) with the Dice ...
Recently, multiresistant Salmonella enterica serovar 1,4,[5],12:i:-, a monophasic variant of S. Typhimurium (1,4,[5],12:i:1,2) emerged, and is now among the most common serovars isolated from humans in many countries. In Greece, monophasic Typhimurium which was recorded for the first time in human isolates in 2007 (0.3% of total isolates), increased sharply thereafter, and since 2009 is the third most frequent serovar. In the present study, 119 S. enterica 1,4,[5],12:i:- strains of human, animal and food origin, isolated during the period between 2006 and 2011, were examined. Strains verified as monophasic Typhimurium variants by polymerase chain reaction (PCR) (97 strains), were further characterised by phenotypic (antibiotic resistance and phage typing) and molecular (pulsed-field gel electrophoresis - PFGE) methods. The results indicate that multiple clones of multiresistant monophasic Typhimurium are circulating in Greece. The most frequently encountered clone in humans and pigs was that of phage
Sequence analysis suggested that these isolates harboured the vanB1 gene. The isolates were susceptible to the majority of antimicrobial agents tested, with the exception of chloramphenicol, erythromycin and vancomycin, and showed distinct profiles of high-level resistance to aminoglycosides. Analysis of the clonal relatedness of the vanB E. faecalis isolates showed similar pulsed-field gel electrophoresis profiles. To our knowledge, this is the first report of the occurrence of enterococcal strains carrying vanB genes in Brazil ...
Abstract: To investigate phage activity in the rumen, a method for quantifying phage has been developed. By differential centrifugation and ultrafiltration, phage particles were separated and concentrated from ruminal fluid. Linear double-stranded DNA from this fraction containing predominantly tailed phage was isolated and separated by size, using pulsed-field gel electrophoresis (PFGE). Laser densitometry of gel photographs allowed the numbers of phages with DNA in each size region to be calculated and, therefore, the total numbers per milliliter of ruminal fluid to be estimated ...
Eleven Salmonella spp. isolates with the antigenic type 11:z41:e,n,z15 - not referred to in the 9th edition of the White-Kauffman-Le Minor Scheme - were identified at the National Reference Laboratory for Salmonella in Greece. Their pulsed-field gel electrophoresis profiles were indistinguishable. No apparent epidemiological link has yet been identified; the results of a case-case study are pending.
In article ,3j1dqu$3t1 at mserv1.dl.ac.uk,, MS WF HONG [GEN]53872 ,GEN282E at ccs1.cc.monash.edu.au, writes: , Dear netters, , I am working on genomic mapping of Pseudomonas by pulse field , gel electrophoresis. At this stage, I have a problem in separating a , 173kb doublet. Does any one have suggestions on how to solve this , problem. , , , Thanks in advance. , , , , W. F. Hong , Well, the best way probably is making a gene library and pulling out linking clones hybridizing the library with the 173kb doublet probe.Making the library shouldnt be problem considering the high and reliable packaging efficiency of commercial pack. mix (Stratagene) like Gigapack Good luck. Goran Biukovic , biukovic at olimp.irb.hr ...
Over past 10 years, Rx Biosciences has designed and developed novel modules and technologies to enhance research in protein engineering and genetic recombination. Staff at Rx Biosciences come from diverse and strong backgrounds often with years of research experience. With a state-of-the-art research facility, Rx Biosciences has a up-to-date infrastructure for handling small and large scale projects. Our labs are well equipped with the latest scientific devices including, fully automated Affymetrix platform for micro-array analysis, ABI, Illumina and Beckman sequencers, Pulse Field Gel Electrophoresis Apparatus, ultra-centrifuge, liquid handling platform, cell culture and animal house facilities.. ...
Early genetic studies showed conservation of gene order in the enteric bacteria. Two recent methods using pulsed field gel electrophoresis (PFGE) to determine the physical map of the genome are: (i) partial digestion with the endonuclease I-CeuI, which digests the DNA of bacteria in the rrn operon f …
DI-fusion, le Dépôt institutionnel numérique de lULB, est loutil de référencementde la production scientifique de lULB.Linterface de recherche DI-fusion permet de consulter les publications des chercheurs de lULB et les thèses qui y ont été défendues.
The SageELF is an electrophoresis system that separates DNA or protein samples by size, and then fractionates the whole sample, or section of sample, into 12 fractions. The system is equipped with pulsed-field electrophoresis for resolving large DNA.. Fractionation ranges are estimated and adjusted in software, and fractions are collected in buffer. One sample is fractionated on a single precast agarose cassette, and one or two cassettes may be processed at one time.. Benefits of the SageELF System: ...
This chapter summarizes the recent findings of bacterial genomics and comments on the themes and trends which are emerging. A variety of techniques and methods are available to construct physical maps, and those most commonly employed involve pulsed field gel electrophoresis (PFGE) of macrorestriction fragments generated by digesting intact genomic DNA, immobilized in agarose plugs, with rare-cutting enzymes. Hybridization techniques are often used to construct a map and to deduce the positions of genetic markers. In recent years significant effort has been devoted to developing direct-mapping techniques for large DNA molecules that do not require gel electrophoresis. Among the more promising of these are two new methods known as DNA combing and optical mapping, both of which make use of fluorescence microscopy and image analysis to visualize single DNA molecules. Overall, bacterial genomes range in size from about 0.6 to 9.4 Mb. In a recent review, it was suggested that there may be a relationship
STROUD WATER RESEARCH CENTER INC - Since 1967, Stroud Water Research Center s scientists and educators have been focused on one thing fresh water. At the heart ...
A total of 486 Listeria monocytogenes isolates originating from 17 Finnish food processing plants (representing meat, poultry, fish, and dairy production) were collected and typed by automated ribotyping using EcoRI as the restriction enzyme. The isolates were divided into 16 different ribotypes (RTs). Some of these isolates (121), representing all EcoRI types and 16 food plants, were subjected to ribotyping with the PvuII enzyme, to pulsed-field gel electrophoresis (PFGE) typing with AscI and SmaI restriction enzymes, and to serotyping with O-antigen antisera. Nineteen ribotypes were generated with PvuII, 42 macrorestriction patterns were generated with AscI and 24 with SmaI, and three serotypes were generated with antisera. When the results were combined, the overall number of RTs was 23, and that of the PFGE types was 46. Thus, the overall discrimination power of PFGE was higher (discrimination index [DI] 0.966) than that of ribotyping (DI 0.906). The most common serotype (90.1% of the isolates) was
In the United States, methicillin-resistant Staphylococcus aureus (MRSA) with the USA300 pulsed-field gel electrophoresis type causes most community-associated MRSA infections and is an increasingly common cause of health care-associated MRSA infections. USA300 probably emerged during the early 1990s. To assess the spatiotemporal diffusion of USA300 MRSA and USA100 MRSA throughout the United States, we systematically reviewed 354 articles for data on 33,543 isolates, of which 8,092 were classified as USA300 and 2,595 as USA100. Using the biomedical literature as a proxy for USA300 prevalence among genotyped MRSA samples, we found that USA300 was isolated during 2000 in several states, including California, Texas, and midwestern states. The geographic mean center of USA300 MRSA then shifted eastward from 2000 to 2013. Analyzing genotyping studies enabled us to track the emergence of a new, successful MRSA type in space and time across the country ...
The aim of the study was to determine antimicrobial resistance and genotypic characteristics of L. monocytogenes isolated from food of animal origin from different parts of Poland during years 2013-2016. A total of 146 isolates were tested using a microbroth dilution method, whereas virulence genes and molecular serogroups were identified by PCR. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) methods were used to analyze the genotypic relationship of the strains. Altogether, 102 pulsotypes grouped into 7 clusters and 24 sequence types, including 3 new types, were identified. Most of the strains clustered into individual patterns were originated from different food products and were isolated in different geographical regions at various time. L. monocytogenes was mostly resistant to oxacilin (90.4% strains), clindamycin (54.1%) and ceftriaxone (49.3%). A multiresistance patterns, mainly to ceftriaxone, oxacillin together with other antimicrobials, were observed among ...
Objective To determine the penicillin resistance and serotype distribution of Streptococcus pneumoniae strains and to identify clonal relationships of isolates resistant to penicillin by means of pulsed-field gel electrophoresis (PFGE). ...
Methicillin-resistant Staphylococcus aureus (MRSA) is a significant pathogen in both human and veterinary medicine. The importance of companion animals as reservoirs of human infections is currently unknown. The companion animals of 49 MRSA-infected outpatients (cases) were screened for MRSA carriage, and their bacterial isolates were compared with those of the infected patients using Pulsed-Field Gel Electrophoresis (PFGE). Rates of MRSA among the companion animals of MRSA-infected patients were compared to rates of MRSA among companion animals of pet guardians attending a
Pulsed field gel electrophoresis is a technique used for the separation of large deoxyribonucleic acid (DNA) molecules by applying to a gel matrix an electric field that periodically changes direction. Standard gel electrophoresis techniques for separation of DNA molecules provided huge advantages for molecular biology research. However, it was unable to separate very large molecules of DNA effectively. DNA molecules larger than 15-20 kb migrating through a gel will essentially move together in a size-independent manner. At Columbia University in 1984, David C. Schwartz and Charles Cantor developed a variation on the standard protocol by introducing an alternating voltage gradient to improve the resolution of larger molecules. This technique became known as pulsed-field gel electrophoresis (PFGE). The development of PFGE expanded the range of resolution for DNA fragments by as much as two orders of magnitude. The procedure for this technique is relatively similar to performing a standard gel ...
in Microbial Drug Resistance : Mechanism, Epidemiology, & Disease (2002), 8(3), 193-200. Stenotrophomonas maltophilia is a nosocomial pathogen with an intrinsic broad-spectrum resistance to beta-lactam compounds and other antibacterial agents. It produces two chromosomal beta-lactamases: a ... [more ▼]. Stenotrophomonas maltophilia is a nosocomial pathogen with an intrinsic broad-spectrum resistance to beta-lactam compounds and other antibacterial agents. It produces two chromosomal beta-lactamases: a clavulanic acid-sensitive class A (L2) and a tetrameric carbapenemase (L1 or BlaS). We screened 40 S. maltophilia multidrug-resistant clinical isolates recovered between 1995 and 1998 in the Varese Hospital (Italy) for the presence of the metallo-beta-lactamase. The isolates were investigated by phenotypic profiling (enzymatic activity and antibiotic resistance pattern) and molecular methods such as PCR and pulsed-field gel electrophoresis (PFGE) to reveal intraspecies diversity. For the tested ...
During 2015-2016, we evaluated the performance of whole-genome sequencing (WGS) as a routine typing tool. Its added value for microbiological and epidemiologic surveillance of listeriosis was compared with that for pulsed-field gel electrophoresis (PFGE), the current standard method. A total of 2,743 Listeria monocytogenes isolates collected as part of routine surveillance were characterized in parallel by PFGE and core genome multilocus sequence typing (cgMLST) extracted from WGS. We investigated PFGE and cgMLST clusters containing human isolates. Discrimination of isolates was significantly higher by cgMLST than by PFGE (p<0.001). cgMLST discriminated unrelated isolates that shared identical PFGE profiles and phylogenetically closely related isolates with distinct PFGE profiles. This procedure also refined epidemiologic investigations to include only phylogenetically closely related isolates, improved source identification, and facilitated epidemiologic investigations, enabling identification
A physical map of the chromosome of Neisseria gonorrhoeae FA1090 has been constructed. Digestion of strain FA1090 DNA with NheI, SpeI, BglII, or PacI resulted in a limited number of fragments that were resolved by contour-clamped homogeneous electric field electrophoresis. The estimated genome size was 2,219 kb. To construct the map, probes corresponding to single-copy chromosomal sequences were used in Southern blots of digested DNA separated on pulsed-field gels, to determine how the fragments from different digests overlapped. Some of the probes represented identified gonococcal genes, whereas others were anonymous cloned fragments of strain FA1090 DNA. By using this approach, a macrorestriction map of the strain FA1090 chromosome was assembled, and the locations of various genetic markers on the map were determined. Once the map was completed, the repeated gene families encoding Opa and pilin proteins were mapped. The 11 opa loci of strain FA1090 were distributed over approximately 60% of ...
Restriction endonuclease patterns generated by Pulsed-Field Gel Electrophoresis (PFGE) were used to compare 96 strains of dairy propionibacteria originating from dairy products, international and indu
This study analyzed 42 strains collected between 2009C2012 from different hospitals in Beyrouth and North Lebanon to raised understand the epidemiology and carbapenem resistance mechanisms in our collection and to compare the robustness of pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST), repetitive sequence-based PCR (rep-PCR) and is an opportunistic gram negative pathogen involved in a wide quantity of nosocomial infections like ventilator-associated pneumonia, bloodstream, urinary tract, wound and meningitis infections frequently associated with a high rate of mortality and morbidity [1]. 18 were international clones and 8 European or Asian restricted clones. The International clone II was the major clone reported in 34 countries in Europe, Asia, Africa Australia, USA, and South America. To track and monitor these outbreaks, denote strain relatedness and assign an outbreak strain to its corresponding clonal lineage, many typing methods with different intrinsic ...
Figure 1. Dendogram derived from PFGE profiles of a) ApaI and b) AscI macrorestriction, showing pattern similarity among the 22 L. monocytogenes strains of the study. Clusters are indicated on the left side of the Figure as well as a 90 % similarity line. PFGE groups are shown in rectangles on the right side of the figure. The source and other information associated with each strain are also shown.. The analysis of macrorestriction patterns obtained by PFGE-ApaI showed 68 % similarity of the 22 strains studied, distributed in 2 clusters: I and II as shown in Figure 1a. Closely related isolates (greater than 90 % similarity in banding patterns) were assigned a PFGE group. Cluster I had 3 isolates and included clone 001 and a ground beef strain (BS78) with a similarity of 83 %. Cluster II had 19 strains and included the PFGE-ApaI group 1 that grouped 15 strains with 92 % similarity. This group contained 80 % (12/15) of the ground beef strains, the strain isolated in the human sporadic case of 2008 ...
Figure 1. Dendogram derived from PFGE profiles of a) ApaI and b) AscI macrorestriction, showing pattern similarity among the 22 L. monocytogenes strains of the study. Clusters are indicated on the left side of the Figure as well as a 90 % similarity line. PFGE groups are shown in rectangles on the right side of the figure. The source and other information associated with each strain are also shown.. The analysis of macrorestriction patterns obtained by PFGE-ApaI showed 68 % similarity of the 22 strains studied, distributed in 2 clusters: I and II as shown in Figure 1a. Closely related isolates (greater than 90 % similarity in banding patterns) were assigned a PFGE group. Cluster I had 3 isolates and included clone 001 and a ground beef strain (BS78) with a similarity of 83 %. Cluster II had 19 strains and included the PFGE-ApaI group 1 that grouped 15 strains with 92 % similarity. This group contained 80 % (12/15) of the ground beef strains, the strain isolated in the human sporadic case of 2008 ...
TY - JOUR. T1 - Insertion element IS3-based PCR method for subtyping Escherichia coli O157. T2 - H7. AU - Thompson, Curt J.. AU - Daly, Claire. AU - Barrett, Timothy J.. AU - Getchell, Jane P.. AU - Gilchrist, Mary J R. AU - Loeffelholz, Mike J.. PY - 1998/5. Y1 - 1998/5. N2 - An Escherichia coli O157:H7 subtyping method based on PCR amplification of variable DNA sequences between the repetitive element IS3 was developed. Template DNA was prepared by boiling cells in Chelex. Two separate IS3 PR amplifications were performed for each isolate: one with a single primer (primer IS3A) and one with two primers (primers IS3A and IS3B). The IS3 PCR subtyping method was applied to 35 epidemiologically related and unrelated E. coli O157:H7 isolates that had been previously characterized by pulsed- field gel electrophoresis (PFGE). PFGE identified 25 different subtypes (difference of one or more bands). PCR with single-primer IS3A and primer pair IS3A-IS3B identified 6 and 14 different subtypes, ...
The RESOLUTION System is based on PathoGenetixs proprietary Genome Sequence Scanning™ (GSS™) technology, which enables pathogen serotype identification and strain typing in just five hours, directly from complex mixtures such as environmental, clinical and enriched food samples. Initially developed to detect bio-threat pathogens in environmental samples under a five-year, $50-million contract through the Department of Homeland Security, the breakthrough GSS technology isolates and analyzes DNA direct from complex mixtures-without the need for a pure culture isolate. The strain type information provided by GSS is comparable in resolution to pulsed field gel electrophoresis (PFGE), one of the current gold standards for pathogen identification ...
Main Article. The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.. ...
Advanced Analytical Technologies Inc AATI has received an RampD 100 Award from RampD Magazine for its FEMTO Pulse automated pulsed-field capillary electrophoresis CE instrument. As the only automated pulsed-field CE system on the market, the FEMTO Pulse won the award for its ability to separate and analyze large nucleic acid fragments span...
Pulsed field gel electrophoresis (PFGE) has provided a very reliable system for separation of DNA fragments greater than 50 kb and has made a significant impact on the analysis of both prokaryotic and eukaryotic genomes. The chapters in this book cover both the theory behind this very important technique and present detailed protocols for many of its major uses.
(2002) Yamaguchi et al. J.Infect.Dis.. A molecular epidemiological analysis was performed to reveal the clonal association of Staphylococcus aureus strains isolated from patients with bullous impetigo. Pulsed-field gel electrophoresis w...
Allele, Antigen, Disease, Electrophoresis, Gene, Identification, Methods, Molecular Typing, Multilocus Sequence Typing, Neisseria, Neisseria Meningitidis, Patients, Pulsed-field Gel Electrophoresis, Strains
The resultant PCR solution was cloned into a TA vector and remodeled into E. coli, which underwent ampicillin variety (a hundred mg/mL) on luria broth plates
In a recent article Epidemiology 1990; 1:421-429 I resurrected some historical criticisms of conventional statistics in non-randomized, non-randomly sampled studies, and suggested some improvements to current practice in response to these criticisms. Here, I propose that some resolution can be achieved by separating data analysis into...
I was officially diagnosed in March 2010. I was dating a man who I thought I would one day marry. I had my first outbreak 3 years into the relationship.
Forty-seven Salmonella Typhimurium (33 zoonotic, 14 clinical) strains were tested for antimicrobial resistance using the standard disk diffusion method. The presence of relevant resistance genes and class 1 integrons were investigated by using PCR. Pulsed-field gel electrophoresis (PFGE) and plasmid profiling were carried out to determine the genomic diversity of Salmonella Typhimurium. Approximately 57.4 of the S. Typhimurium strains were multidrug resistant (MDR) and showed high resistance rates to tetracycline (70.2), sulfonamides (57.4), streptomycin (53.1), ampicillin (29.7), nalidixic acid (27.6), kanamycin (23.4), chloramphenicol (21.2), and trimethoprim (19.1). Resistance towards cephalosporins was noted for cephalothin (27.6), cephradine (21.2), amoxicillin clavulanic acid (17.0), and cephalexin (17.0). Resistance genes, bla(TEM), strA, aadA, sul1, sul2, tetA, tetB, and tetC, were detected among the drug-resistant strains. Thirty-three strains (70.2) carried class 1 integrons, which ...
This study addresses the epidemiologic relatedness of a collection of Pseudomonas aeruginosa isolates from cystic fibrosis patients attending the Pediatric Clinic, Catania, Sicily. Genome macrorestriction analysis after pulsed field gel electrophoresis (PFGE) was used to characterise all strains. Furthermore, a rapid typing procedure, developed in this study, based on polymerase chain reaction amplified ribosomal DNA spacer polymorphisms (PCR-ribotyping), straight from bacterial cultures, was used. On the basis of macrorestriction analysis after PFGE, persistence of infection was shown in all patients; two cross-transmission episodes were identified in the nosocomial as well as in the familiar environment. PCR-ribotyping proved to be useful for a DNA-based identification test, suitable for screening purposes. The rapid amplification protocol here tested is proposed to evaluate the discriminatory power of other specific target sequences in PCR-based typing assays, for epidemiologic purposes.
BACKGROUND: Avoparcin, cross-resistance with vancomycin, was added as feed-additive since 1970s and was prohibited in 1997 in Korea. After avoparcin was banned we examined prevalence and genetic relatedness of VRE in enterococci isolated from livestock and humans. MATERIALS AND METHODS: Using enrichment broth and 6 microgram/mL vancomycin-containing enterococcosel selective agar, vancomycin-resistant enterococci (VRE) were isolated from fecal sample of 255 pigs of 8 farms, 431 chickens of 9 farms, and 328 humans (Food industry employee and Institution cafeteria employee) of 5 public health centers, and 100 raw chicken meats from April to June 2003. Antimicrobial susceptibility was examined by disk diffusion and minimum inhibitory concentrations (MICs), and E-test. Species identification and genotyping were done by multiplex PCR method. Pulsed-field gel electrophoresis (PFGE) of vanA-type VRE isolates was performed by CHEF-Mapper system. RESULTS: 19 isolates from 255 pigs, 122 isolates from 431 ...
Klebsiella oxytoca is primarily a health care-associated pathogen acquired from environmental sources. During October 2006-March 2011, a total of 66 patients in a hospital in Toronto, Ontario, Canada, acquired class A extended-spectrum β-lactamase-producing K. oxytoca with 1 of 2 related pulsed-field gel electrophoresis patterns. New cases continued to occur despite reinforcement of infection control practices, prevalence screening, and contact precautions for colonized/infected patients. Cultures from handwashing sinks in the intensive care unit yielded K. oxytoca with identical pulsed-field gel electrophoresis patterns to cultures from the clinical cases. No infections occurred after implementation of sink cleaning 3×/day, sink drain modifications, and an antimicrobial stewardship program. In contrast, a cluster of 4 patients infected with K. oxytoca in a geographically distant medical ward without contaminated sinks was contained with implementation of active screening and contact ...
Active surveillance of Vibrio parahaemolyticus infection among hospitalized patients in Calcutta, India, showed the appearance of the O4[ratio ]K68 serovar for the first time in March 1998 alongside the continued predominant incidence of the O3[ratio ]K6 serovar. Strains belonging to both these serovars have been reported to possess pandemic potential. The genomes of O3[ratio ]K6 and O4[ratio ]K68 strains and for comparison, non-O3[ratio ]K6 and non-O4[ratio ]K68 strains isolated from two different countries, India and Thailand, were examined by different molecular techniques to determine their relatedness. The O3[ratio ]K6 and O4[ratio ]K68 strains from Calcutta and Bangkok carried the tdh gene but not the trh gene. Characterization of representative strains of these two serovars by ribotyping and by arbitrarily primed-polymerase chain reaction (AP-PCR) showed that the isolates had identical ribotype and DNA fingerprint. Pulsed-field gel electrophoresis (PFGE) performed with the same set of ...
HANIFAH, YASMIN BINTI ABU (2009) Resistant Phenotypes and Genetic Diversity of Nosocomial Acinetobacter baumanii Using Pulsed-Field Gel Electrophoresis in Intensive Care Unit, University Malaya Medical Centre. Full text not available from this repository ...
To increase the utility of the previously constructed physical map of the chromosome of Neisseria gonorrhoeae FA1090, 28 additional genetic markers were localized on the map. Cloned gonococcal genes were used to probe Southern blots of restriction enzyme-digested DNA separated on pulsed-field gels, thus identifying the fragment in each of several digests to which the probe hybridized and the map location of each gene. The addition of the new markers brings the total number of mapped loci for this strain to 68; the locations of all of those markers on the updated map are shown. ...
Boey, C.M. (2010) Reflections on the significance of the relationship between mind and body in medicine. Journal of Health and Translational Medicine, 13 (1). pp. 3-11.. Koh, P.S.; Cha, K.H.; Lucy, C.; Rampal, S.; Yoong, B.K. (2012) Superior somatic pain relief and improved visceral pain control is achieved using pre-emptive analgesia for laparoscopic cholecystectomy: A randomized controlled trial. Journal of Health and Translational Medicine, 15 (2). pp. 1-7. ISSN 1823-7339. Thong, K.L.; Puthucheary, S.D.; Boey, C.C.M.; Pang, T. (1999) Investigation of a recurrent case of Salmonellosis due to Salmonella bovismorbificans and Salmonella matopeni using pulsed-field gel electrophoresis analysis and antibiograms. Journal of Health and Translational Medicine, 4 (2). pp. 104-109. ISSN 1823-7339. ...
Since the 2006 discovery of the Acinetobacter baumannii strain AYE AbaR1 resistance island, similar elements have been reported in numerous members of this species. As AbaR1 is distantly related to Tn7, we have renamed it TnAbaR1. TnAbaR transposons are known to carry multiple antibiotic resistance- and efflux-associated genes, although none have been experimentally studied en bloc. We deleted the TnAbaR transposon in A. baumannii A424, which we have designated TnAbaR23, and characterized independent deletion mutants DCO163 and DCO174. The NotI pulsed-field gel electrophoresis (PFGE) profile of strain DCO174 was consistent with targeted deletion of TnAbaR23 alone, but strain DCO163 apparently harbored a second large genomic deletion. Nevertheless, subtractive amplification targeting 52 TnAbaR and/or resistance-associated loci yielded identical results for both mutants and highlighted genes lost relative to strain A424. PCR mapping and genome sequencing revealed the entire 48.3-kb sequence of ...
Electrophoresis, Human, Meat, Meat Products, Methicillin, Methicillin-resistant, Mrsa, Multidrug Resistance, Multilocus Sequence Typing, Pulsed-field Gel Electrophoresis, Staphylococcus, Staphylococcus Aureus
Contributors. Forword.. Preface.. Dedication.. I. Introduction to Healthcare Associated Infections and Their Control.. 1. The Hospital and Ambulatory Care Environment (Anne Y. Chen and Hiren Pokharma).. 2. Pathogen Transmission in the Healthcare Setting (Sonja Hansen and Ralf-Peter Vonberg).. 3. Infection Control Basics (Louise-Marie Dembry and Carlos Torres-Viera).. 4. Cost-Effectiveness of IC Program (Marc-Oliver Wright and Eli N. Perencevich).. 5. Outbreak Investigations (Importance of the Healthcare Epidemiologist) (Marcus J. Zervos).. 6. Pathogen Elimination: Antibiotic and Disinfectant Use and the Development of Resistance (Steven L. Foley, Beilei Ge, Carl M. Schroeder, and Arron M. Lynne).. II. Techniques to Characterize Nosocomial Pathogens.. 7. Rapid PCR Screening Methods (Ngolela Esther Babady, Frankling Cockerill and Robin Patel).. 8. Restriction Analysis Techniques (Richard V. Goering, Mary Stemper, SanjayShukla and Steven Foley).. 9. Pulsed-field Gel Electrophoresis (Mary Stemper, ...
With the fingerprint data module, BioNumerics offers the most comprehensive tools for processing electrophoresis profiles and spectra.