Application of Random Amplified Polymorphic DNA (RAPD) and Pulsed-Field Gel Electrophoresis (PFGE) Analysis toListeriamonocytogenes: A Review of Methodology and Results

2003 ◽  
Vol 17 (3) ◽  
pp. 217-236 ◽  
Author(s):  
Robert E. Levin
Keyword(s):  
Gel Electrophoresis ◽  
Random Amplified Polymorphic Dna ◽  
Pulsed Field Gel Electrophoresis ◽  
Pfge Analysis ◽  
Pulsed Field

scholarly journals Coagulase gene variants associated with distinct populations of Staphylococcus aureus

2003 ◽  
Vol 130 (2) ◽  
pp. 207-219 ◽  
Author(s):  
P. E. CARTER ◽  
K. BEGBIE ◽  
F. M. THOMSON-CARTER
Keyword(s):  
Staphylococcus Aureus ◽  
Gel Electrophoresis ◽  
Multilocus Sequence Typing ◽  
Clonal Complex ◽  
Protein A ◽  
Pulsed Field Gel Electrophoresis ◽  
Gene Variants ◽  
Pfge Analysis ◽  
Pulsed Field ◽  
Clonal Distribution

An identifying characteristic of Staphylococcus aureus is the production of staphylocoagulase (coagulase). The aim of this study was to determine the clonal distribution of coagulase gene (coa) variants within populations of S. aureus defined by multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and protein A variation. The N-terminal region of the coa gene from 43 methicillin-susceptible (MSSA) and 252 methicillin-resistant (MRSA) S. aureus human isolates and 9 animal S. aureus isolates was amplified and digested with HinfI. Twelve types were identified amongst the MSSA isolates and the majority (93%) of MRSA isolates were assigned to 5 of the 12 types. MLST and PFGE analysis identified epidemic populations of MRSA and each epidemic population was characterized by a different coagulase type. Nine of the 12 MLST-defined clonal complex ancestral genotypes recently described each carried a different coagulase type suggesting that coagulase evolution and the evolution of the clonal complexes are intimately related.

scholarly journals Subtyping of Haemophilus influenzaeStrains by Pulsed-Field Gel Electrophoresis

1999 ◽  
Vol 37 (7) ◽  
pp. 2142-2147 ◽  
Author(s):  
Mitsumasa Saito ◽  
Akiko Umeda ◽  
Shin-ichi Yoshida
Keyword(s):  
Cerebrospinal Fluid ◽  
Haemophilus Influenzae ◽  
Gel Electrophoresis ◽  
Invasive Disease ◽  
Pulsed Field Gel Electrophoresis ◽  
Pfge Analysis ◽  
Pulsed Field ◽  
Pfge Typing ◽  
Household Contacts ◽  
Serotype B

A total of 200 isolates of Haemophilus influenzae were analyzed by serotyping, biotyping, and pulsed-field gel electrophoresis (PFGE). A total of 178 epidemiologically unrelated strains of H. influenzae demonstrated a variety of genome patterns by PFGE, and 165 genotypes were thus obtained in this study. PFGE typing proved to have a much stronger discriminatory power than either serotyping or biotyping. Six serotype b strains were all classified into discrete genotypes. A PFGE analysis of 18 strains obtained from the nasopharynx, blood, and cerebrospinal fluid of patients with meningitis also supported the hypothesis that invasive H. influenzaedisseminates from the nasopharynx to the bloodstream and then subsequently to other body sites. PFGE typing of 10 other strains isolated from household contacts of patients with H. influenzae infection revealed that the strain that caused theH. influenzae infection often colonized the nasopharynges of household contacts. Our findings suggest that PFGE analysis is useful for the epidemiological study of H. influenzaeinfection, even when the invasive disease is caused by serotype b strains.

scholarly journals Prevalence and Dissemination of oqxAB in Escherichia coli Isolates from Animals, Farmworkers, and the Environment

2010 ◽  
Vol 54 (10) ◽  
pp. 4219-4224 ◽  
Author(s):  
Jingjing Zhao ◽  
Zhangliu Chen ◽  
Sheng Chen ◽  
Yuting Deng ◽  
Yahong Liu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Quinolone Resistance ◽  
Pfge Analysis ◽  
Pulsed Field ◽  
E Coli ◽  
Animal Farms ◽  
Food Animals

ABSTRACT OqxAB has recently been identified as one of the mechanisms of plasmid-mediated quinolone resistance (PMQR). Compared to what is observed for other PMQR determinants, there is a paucity of data with regard to the prevalence and epidemiology of OqxAB and its contribution to resistance to different antimicrobials. In this study, the prevalence and dissemination of oqxAB and other PMQR genes in Escherichia coli isolates from animals, farmworkers, and the environment in 2002 in China were investigated. Of the 172 E. coli isolates, 39.0% carried oqxA, while only 4.1%, 2.9%, and 0.6% carried qnr (1 qnrB6 isolate, 5 qnrS1 isolates, and 1 qnrD isolate), qepA, and aac(6′)-Ib- cr, respectively. Among the 33 isolates from farmworkers, 10 (30.3%) were positive for oqxA. oqxAB was associated with IS26 and was carried on the 43- to 115-kb IncF transferable plasmid. Transconjugants carrying oqxAB showed 4- to 16-fold increases in the MICs of quinolones, 16- to 64-fold increases in the MICs of quinoxalines, 8- to 32-fold increases in the MICs of chloramphenicol and trimethoprim-sulfamethoxazole, and 4- to 8-fold increases in the MICs of florfenicol compared to the levels for the recipient. The pulsed-field gel electrophoresis (PFGE) analysis showed that the high levels of prevalence and dissemination of oqxA B in E. coli in animal farms were primarily due to the transmission of plasmids carrying oqxAB, although clonal transmission between human and swine E. coli isolates was observed. It is concluded that oqxAB was widespread in animal farms in China, which may be due to the overuse of quinoxalines in animals. This study warrants the prudent use of quinoxalines in food animals.

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.

scholarly journals Molecular epidemiology of fluoroquinolone-resistant Escherichia coli bloodstream isolates from patients admitted to European cancer centers.

1996 ◽  
Vol 40 (2) ◽  
pp. 387-392 ◽  
Author(s):  
M Oethinger ◽  
S Conrad ◽  
K Kaifel ◽  
A Cometta ◽  
J Bille ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cancer Patients ◽  
Gel Electrophoresis ◽  
Random Amplified Polymorphic Dna ◽  
Pulsed Field Gel Electrophoresis ◽  
Fluoroquinolone Resistance ◽  
Chromosomal Dna ◽  
Cancer Centers ◽  
Pulsed Field ◽  
Time Period

Previous reports have suggested an increasing incidence of highly fluoroquinolone-resistant Escherichia coli causing bacteremia among cancer patients on prophylactic therapy. We used genotyping by pulsed-field gel electrophoresis of chromosomal DNA digests and random amplified polymorphic DNA fingerprinting to study clonal relationships among such isolates obtained at 10 cancer centers located across Europe and the Middle East. Analysis by both methods indicated that isolates from different centers were genotypically unrelated to each other. There were five centers from which more than one individual patient isolate was available, and most demonstrated significant within-center genetic diversity of strains. Strains shared among patients could be identified at two centers. At the center with the largest number of bloodstream isolates from cancer patients available, fluoroquinolone-resistant control isolates from surgical patients and fluoroquinolone-susceptible control isolates from patients admitted to medical services during the same time period were unrelated to resistant cancer patient isolates and to each other as well. A substantial number of fluoroquinolone-resistant isolates (19 of 58) were nontypeable by pulsed-field gel electrophoresis. Fluoroquinolone resistance was commonly associated with multiple antibiotic resistance to chemically unrelated antibacterial agents irrespective of the origin of the isolates.

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