scholarly journals Pulsed-Field Gel Electrophoresis (PFGE) Pattern Analysis and Chlorine-Resistance of Legionella pneumophila Isolated from Hot Spring Water Samples

2014 ◽  
Vol 19 (1) ◽  
pp. 33-38 ◽  
Author(s):  
KATSUNORI FURUHATA ◽  
NAOTO ISHIZAKI ◽  
NAO UMEKAWA ◽  
MIYUKI NISHIZIMA ◽  
MASAFUMI FUKUYAMA
Keyword(s):  
Legionella Pneumophila ◽  
Water Samples ◽  
Gel Electrophoresis ◽  
Pattern Analysis ◽  
Hot Spring ◽  
Pfge Pattern ◽  
Spring Water ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field

scholarly journals Genetic diversity of atypical Aeromonas salmonicida studied by pulsed-field gel electrophoresis

1999 ◽  
Vol 123 (2) ◽  
pp. 299-307 ◽  
Author(s):  
M.-L. HÄNNINEN ◽  
V. HIRVELÄ-KOSKI
Keyword(s):  
Genetic Heterogeneity ◽  
Gel Electrophoresis ◽  
Pattern Analysis ◽  
Genetic Relatedness ◽  
Profile Analysis ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Aeromonas Salmonicida ◽  
Fish Farms ◽  
Pulsed Field

Pulsed-field gel electrophoresis (PFGE) pattern analysis with XbaI restriction enzyme was used to study the genetic heterogeneity of 88 atypical Aeromonas salmonicida strains which were earlier or during this study characterized phenotypically, by ribotyping (ClaI/PstI) and by plasmid profile analysis. The strains of certain ribotypes were also analysed by digestion with SpeI. The strains represented different geographic locations: Finland (72 strains), Iceland (5 strains), Norway (5 strains), Sweden (4 strains) and Denmark (2 strains), and they were from 17 fish species during 1981–97. Thirty-one PFGE genotypes found among these strains correlated well with the ribotypes, and in most cases PFGE pattern analysis subdivided ribotypes into several PFGE genotypes, and further within a PFGE genotype into subtypes. XbaI and SpeI digests produced concordant results. In most cases, PFGE patterns of strains with the same ribotype shared many fragments, suggesting genetic relatedness. PFGE patterns of most Norwegian and Icelandic strains isolated during an approximately 10-year period had the same ribotype and their PFGE patterns shared most fragments, suggesting close genetic relatedness. Moreover, atypical strains of ribotypes B/B and H/H isolated from the same Finnish fish farms had closely related patterns suggesting genetic stability and persistence of these genotypes. Genotype 29 of Achromogenic strains was strongly associated with disease of Finnish arctic char and grayling. PFGE was shown to be a distinguishing method to study the genetic heterogeneity of atypical A. salmonicida. This method is applicable to studies of the epidemiology of these infections.

scholarly journals Analysis of Legionella pneumophilaSerogroup 1 Isolates by Pulsed-Field Gel Electrophoresis

1999 ◽  
Vol 37 (1) ◽  
pp. 251-254 ◽  
Author(s):  
Thomas J. Marrie ◽  
Shaun Tyler ◽  
Gregory Bezanson ◽  
Charles Dendy ◽  
Wendy Johnson
Keyword(s):  
Nova Scotia ◽  
Legionella Pneumophila ◽  
Gel Electrophoresis ◽  
Genomic Dna ◽  
Plasmid Profile ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Plasmid Content ◽  
Pulsed Field

At a hospital in Halifax, Nova Scotia, Canada, three strains of Legionella pneumophila were detectable based on plasmid content, while the isolates collected at another hospital in Halifax had no plasmids. Genomic DNA was digested withBssHII, SalI, and SpeI and subjected to pulsed-field gel electrophoresis (PFGE). We found no relationship between plasmid profile and PFGE pattern.

scholarly journals Application of pulsed-field gel electrophoresis to the epidemiological characterization ofStaphylococcus intermediusimplicated in a food-related outbreak

1994 ◽  
Vol 113 (1) ◽  
pp. 75-81 ◽  
Author(s):  
F. M. Khambaty ◽  
R. W. Bennett ◽  
D. B. Shah
Keyword(s):  
Gel Electrophoresis ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Western United States ◽  
Single Strain ◽  
Pulsed Field ◽  
Definitive Evidence ◽  
Staphylococcus Intermedius ◽  
Epidemiological Characterization

SUMMARYAn outbreak of food intoxication involving over 265 cases in western United States occurred in October 1991.Staphylococcus intermediuswas implicated as the aetiologic agent. Representative outbreak isolates (five clinical and ten from foods) produced type A enterotoxin. DNA fragments generated by four restriction endonucleases and analysed by pulsed-field gel electrophoresis (PFGE) provided definitive evidence that all isolates from nine different counties in California and Nevada were derived from a single strain. The PFGE pattern of these outbreak isolates was distinct from those of a heterogeneous collection of sevenS. intermediusstrains of veterinary origin and five unrelatedS. aureuslaboratory strains. The data show a significant PFGE pattern heterogeneity not only among members of differentStaphylococcusspecies but also within members of the same species and even the same enterotoxin type. The results indicate that PFGE is a valuable strain-specific discriminator for the epidemiological characterization ofS. intermedius. To our knowledge, this represents the first documented foodborne outbreak caused byS. intermedius. These findings suggest that the presence ofS. intermediusand other species such asS. hyicusin food should be reason for concern.

scholarly journals Optimization of Pulsed-Field Gel Electrophoresis for Legionella pneumophila Subtyping

2010 ◽  
Vol 76 (5) ◽  
pp. 1334-1340 ◽  
Author(s):  
Haijian Zhou ◽  
Hongyu Ren ◽  
Bingqing Zhu ◽  
Biao Kan ◽  
Jianguo Xu ◽  
...  
Keyword(s):  
Legionella Pneumophila ◽  
Gel Electrophoresis ◽  
Water System ◽  
Epidemiological Data ◽  
Pulsed Field Gel Electrophoresis ◽  
Discriminatory Power ◽  
Similarity Coefficients ◽  
Pulsed Field ◽  
Software Analysis ◽  
High Degree

ABSTRACT A total of 32 strains of Legionella pneumophila were used to optimize pulsed-field gel electrophoresis (PFGE) for subtyping of L. pneumophila. Twenty-six isolates of L. pneumophila with various origins and 11 isolates from five different water systems were used as the panels. For optimization of electrophoretic parameters (EPs) of SfiI PFGE, 26 isolates were analyzed with SfiI digestion, using four EPs yielding the same D value. The EP of a switch time of 5 to 50 s for 21 h had the smallest similarity coefficients and was declared the optimal EP for SfiI PFGE of L. pneumophila. By software analysis and pilot study, AscI was chosen as another PFGE enzyme. AscI PFGE could cluster the isolates from each water system into the same or very similar patterns and had a high degree of typing concordance with other molecular methods. In evaluating the discriminatory power of AscI with the panel of 26 isolates, AscI PFGE gave one single pattern and a D value of 100%. AscI PFGE had a high discriminatory power and a high degree of consistency with epidemiological data and other molecular typing methods for L. pneumophila subtyping, and hence, AscI could be used as a restriction enzyme in PFGE subtyping of L. pneumophila.

Monitoring of Chicken Houses and an Attached Egg-Processing Facility in a Laying Farm for Salmonella Contamination between 1994 and 1998

2001 ◽  
Vol 64 (12) ◽  
pp. 1912-1916 ◽  
Author(s):  
TOSHIYUKI MURASE ◽  
KAZUKO SENJYU ◽  
TAKESHI MAEDA ◽  
MASAYUKI TANAKA ◽  
HIROSHI SAKAE ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Salmonella Enterica ◽  
Environmental Samples ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Drain Water ◽  
Pulsed Field ◽  
Processing Facility ◽  
Single Clone

Two chicken houses and an attached egg-processing facility in a laying farm were sampled between 1994 and 1998 to investigate Salmonella contamination. Each of the houses was environmentally controlled and fitted with egg belts that transported eggs from the houses to the egg-processing facility. Four hundred twenty-eight Salmonella isolates were obtained from 904 environmental samples collected from the houses. Two hundred fifty-two of the 428 (58.9%) isolates yielded five serotypes as follows: Salmonella enterica subsp. enterica serovar Livingstone, Salmonella serovar Cerro, Salmonella serovar Montevideo, Salmonella serovar Mbandaka, and Salmonella serovar Corvallis. The remaining (41.1%, 176 of 428) isolates included four other serotypes and isolates that were untypeable. Salmonella isolates obtained from the drain water collected after the washing of the eggs in the egg-processing facility yielded the same serotypes as those found in the chicken houses. Strains having an identical pulsed-field gel electrophoresis (PFGE) pattern were continually recovered from a house for more than 1 year. Several strains of Salmonella Cerro, Salmonella Mbandaka, and Salmonella Montevideo obtained from both the houses and from the egg-processing facility were indistinguishable by PFGE, respectively. These results suggest that Salmonella organisms originating from a single clone colonized the chicken houses and that the egg belts are likely to be one of the means by which Salmonella organisms are spread from one house to the others.

Outbreak of PER-1 and diversity of β-lactamases among ceftazidime-resistant Pseudomonas aeruginosa clinical isolates

2014 ◽  
Vol 63 (3) ◽  
pp. 386-392 ◽  
Author(s):  
Yun Qing ◽  
Kai-Yuan Cao ◽  
Zhi-Li Fang ◽  
Ying-Min Huang ◽  
Xue-Fei Zhang ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Gel Electrophoresis ◽  
Southern China ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Clinical Isolates ◽  
Pulsed Field ◽  
High Degree ◽  
Esbl Producers ◽  
M 14

A growing number of β-lactamases have been reported in Pseudomonas aeruginosa clinical isolates. The aim of this study was to investigate the diversity of β-lactamases in the collection of 51 ceftazidime-resistant P. aeruginosa clinical isolates in four hospitals of southern China. Among these isolates, variable degrees of resistance to other β-lactam and non-β-lactam agents were observed. Pulsed-field gel electrophoresis (PFGE) revealed a high degree of clonality with five main genotypes. Of the 51 isolates tested, 35 (68.6 %) were identified as extended-spectrum β-lactamase (ESBL) producers, with 35 producing PER-1, 1 CTX-M-3, 7 CTX-M-15 and 1 CTX-M-14. Most (82.9 %, 29/35) PER-1-producing isolates were collected from two hospitals between January and April in 2008 and belonged to the same PFGE pattern (pattern B) with similar antibiogram and β-lactamase profiles, which suggested an outbreak of this clone at the time. The prevalence of CTX-M-type ESBL (17.6 %, 9/51) was unexpectedly high. One isolate was identified as producing VIM-2. Furthermore, we also reported an occurrence of a novel OXA-10 variant, OXA-246, in 14 P. aeruginosa isolates. In addition, AmpC overproduction was found to be the β-lactamase-mediated mechanism responsible for ceftazidime resistance in 6 isolates (11.8 %). Our results revealed an overall diversity of β-lactamases and outbreak of a PER-1-producing clone among ceftazidime-resistant P. aeruginosa in southern China.

scholarly journals A Whole-Genome Single Nucleotide Polymorphism-Based Approach To Trace and Identify Outbreaks Linked to a Common Salmonella enterica subsp. enterica Serovar Montevideo Pulsed-Field Gel Electrophoresis Type

2011 ◽  
Vol 77 (24) ◽  
pp. 8648-8655 ◽  
Author(s):  
Henk C. den Bakker ◽  
Andrea I. Moreno Switt ◽  
Craig A. Cummings ◽  
Karin Hoelzer ◽  
Lovorka Degoricija ◽  
...  
Keyword(s):  
Single Nucleotide Polymorphism ◽  
Gel Electrophoresis ◽  
Salmonella Enterica ◽  
Pfge Pattern ◽  
Pulsed Field Gel Electrophoresis ◽  
Whole Genome ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Content Type ◽  
Pulsed Field

ABSTRACTIn this study, we report a whole-genome single nucleotide polymorphism (SNP)-based evolutionary approach to study the epidemiology of a multistate outbreak ofSalmonella entericasubsp.entericaserovar Montevideo. This outbreak included 272 cases that occurred in 44 states between July 2009 and April 2010. A case-control study linked the consumption of salami made with contaminated black and red pepper to the outbreak. We sequenced, on the SOLiD System, 47 isolates with XbaI PFGE pattern JIXX01.0011, a common pulsed-field gel electrophoresis (PFGE) pattern associated with isolates from the outbreak. These isolates represented 20 isolates collected from human sources during the period of the outbreak and 27 control isolates collected from human, food, animal, and environmental sources before the outbreak. Based on 253 high-confidence SNPs, we were able to reconstruct a tip-dated molecular clock phylogeny of the isolates and to assign four human isolates to the actual outbreak. We developed an SNP typing assay to rapidly discriminate between outbreak-related cases and non-outbreak-related cases and tested this assay on an extended panel of 112 isolates. These results suggest that only a very small percentage of the human isolates with the outbreak PFGE pattern and obtained during the outbreak period could be attributed to the actual pepper-related outbreak (20%), while the majority (80%) of the putative cases represented background cases. This study demonstrates that next-generation-based SNP typing provides the resolution and accuracy needed for outbreak investigations of food-borne pathogens that cannot be distinguished by currently used subtyping methods.

scholarly journals Comparative Analysis of Infrequent-Restriction-Site PCR and Pulsed-Field Gel Electrophoresis for Epidemiological Typing ofLegionella pneumophila Serogroup 1 Strains

1998 ◽  
Vol 36 (1) ◽  
pp. 161-167 ◽  
Author(s):  
Serge Riffard ◽  
François Lo Presti ◽  
François Vandenesch ◽  
Françoise Forey ◽  
Monique Reyrolle ◽  
...  
Keyword(s):  
Legionella Pneumophila ◽  
Gel Electrophoresis ◽  
Restriction Site ◽  
Pulsed Field Gel Electrophoresis ◽  
Pfge Analysis ◽  
Pcr Assay ◽  
Pulsed Field ◽  
Restriction Sites ◽  
Dna Restriction ◽  
High Level

Two methods were compared for the analysis of 48 unrelated and epidemiologically related Legionella pneumophila serogroup 1 isolates. These are the infrequent-restriction-site PCR (IRS-PCR) assay with adapters designed for XbaI and PstI restriction sites and the pulsed-field gel electrophoresis (PFGE) analysis determined after DNA restriction with SfiI. Both methods demonstrated a high level of discrimination with a similar capacity for differentiating 23 of the 24 unrelated isolates. PFGE analysis and IRS-PCR assay were both able to identify epidemiologically related isolates of L. pneumophila from three outbreaks. Hence, IRS-PCR assay appears to be a reproducible (intergel reproducibility, 100%) and discriminative (discriminatory index, ≥0.996) tool for typing of Legionella. Compared to PFGE, however, IRS-PCR presented an advantage through ease of performance and with attributes of rapidity and sensitivity of target DNA.

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