Suitability of the Molecular Subtyping Methods Intergenic Spacer Region, Direct Genome Restriction Analysis, and Pulsed-Field Gel Electrophoresis for Clinical and EnvironmentalVibrio parahaemolyticusIsolates

2014 ◽  
Vol 11 (7) ◽  
pp. 520-528 ◽  
Author(s):  
Catharina H.M. Lüdeke ◽  
Markus Fischer ◽  
Patti LaFon ◽  
Kara Cooper ◽  
Jessica L. Jones
Keyword(s):  
Gel Electrophoresis ◽  
Restriction Analysis ◽  
Intergenic Spacer ◽  
Pulsed Field Gel Electrophoresis ◽  
Molecular Subtyping ◽  
Pulsed Field ◽  
Intergenic Spacer Region

scholarly journals The Aquatic Environment as a Reservoir ofVibrio choleraeO1 in Hydrographic Basins of the State of Pernambuco, Brazil

2013 ◽  
Vol 2013 ◽  
pp. 1-5
Author(s):  
Carina Lucena Mendes-Marques ◽  
Vladimir da Mota Silveira Filho ◽  
Ana Paula Rocha da Costa ◽  
Mariana de Lira Nunes ◽  
Sandoval Vieira da Silva Filho ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Genetic Similarity ◽  
Virulence Genes ◽  
Intergenic Spacer ◽  
Pulsed Field Gel Electrophoresis ◽  
N Gene ◽  
Intergenic Spacer Region ◽  
Quiescent Period ◽  
Pathogenic Genes ◽  
Culture Negative

After the worldwide cholera epidemic in 1993, permanent environmental monitoring of hydrographic basins was established in Pernambuco, Brazil, where cholera is endemic. After a quiescent period, 4rfbN (serogroup O1) positive water samples that were culture negative were detected by multiplex single-tube nested PCR (MSTNPCR); 2 of these were alsoctxA (cholera toxin) positive. From May to June 2012, 30V. choleraeO1 isolates were obtained by culturing samples. These isolates were analyzed for the presence of virulence genes by PCR, intergenic spacer region 16S-23S PCR (ISR-PCR), and pulsed field gel electrophoresis (PFGE). The isolates were positive for therfbN gene and negative for the assessed pathogenic genes and were classified into 2 groups by ISR and the same profile by PFGE. Close genetic similarity was observed between them (2012) and environmental strains from 2004 to 2005, indicating the permanence of endemicV. choleraeO1 in the region.

scholarly journals Molecular Subtyping of Clostridium perfringens by Pulsed-Field Gel Electrophoresis To Facilitate Food-Borne-Disease Outbreak Investigations

1999 ◽  
Vol 37 (7) ◽  
pp. 2209-2214 ◽  
Author(s):  
Susan E. Maslanka ◽  
Jared G. Kerr ◽  
Glen Williams ◽  
James M. Barbaree ◽  
Loretta A. Carson ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Gel Electrophoresis ◽  
Disease Outbreaks ◽  
Pulsed Field Gel Electrophoresis ◽  
Single Individual ◽  
Molecular Subtyping ◽  
Pulsed Field ◽  
Food Borne ◽  
Outbreak Investigations ◽  
Food Borne Illness

Clostridium perfringens is a common cause of food-borne illness. The illness is characterized by profuse diarrhea and acute abdominal pain. Since the illness is usually self-limiting, many cases are undiagnosed and/or not reported. Investigations are often pursued after an outbreak involving large numbers of people in institutions, at restaurants, or at catered meals. Serotyping has been used in the past to assist epidemiologic investigations of C. perfringensoutbreaks. However, serotyping reagents are not widely available, and many isolates are often untypeable with existing reagents. We developed a pulsed-field gel electrophoresis (PFGE) method for molecular subtyping of C. perfringens isolates to aid in epidemiologic investigations of food-borne outbreaks. Six restriction endonucleases (SmaI, ApaI, FspI,MluI, KspI, and XbaI) were evaluated with a select panel of C. perfringens strains.SmaI was chosen for further studies because it produced 11 to 13 well-distributed bands of 40 to ∼1,100 kb which provided good discrimination between isolates. Seventeen distinct patterns were obtained with 62 isolates from seven outbreak investigations or control strains. In general, multiple isolates from a single individual had indistinguishable PFGE patterns. Epidemiologically unrelated isolates (outbreak or control strains) had unique patterns; isolates from different individuals within an outbreak had similar, if not identical, patterns. PFGE identifies clonal relationships of isolates which will assist epidemiologic investigations of food-borne-disease outbreaks caused by C. perfringens.

scholarly journals Validity of SmaI-defined genotypes of Campylobacter jejuni examined by SalI, KpnI, and BamHI polymorphisms: evidence of identical clones infecting humans, poultry, and cattle

1998 ◽  
Vol 120 (3) ◽  
pp. 231-237 ◽  
Author(s):  
S. L. W. ON ◽  
E. M. NIELSEN ◽  
J. ENGBERG ◽  
M. MADSEN
Keyword(s):  
Campylobacter Jejuni ◽  
Gel Electrophoresis ◽  
Restriction Site ◽  
Epidemiological Studies ◽  
Pulsed Field Gel Electrophoresis ◽  
Genetic Differences ◽  
Genetic Fingerprinting ◽  
Molecular Subtyping ◽  
Pulsed Field ◽  
Heat Stable

We describe here an examination of the validity of molecular types of Campylobacter jejuni as defined by separation of SmaI-digested DNA using pulsed-field gel electrophoresis (PFGE), recently suggested as part of a molecular subtyping scheme. Thirty-four Danish strains from humans, water, poultry and cattle were assigned to one of six SmaI ‘profile groups’ (PGs), with two additional strains included as genotypically distinct controls. The interstrain relationships were reexamined by PFGE of SalI, KpnI and BamHI-digested DNA, and also by serotyping with heat-stable antigens. All outbreak-related strains were indistinguishable by all criteria, as were two sets of two randomly-isolated human strains. Two groups of indistinguishable isolates contained randomly isolated strains from more than one source (poultry, humans and/or cattle), a finding with significant epidemiological connotations. All ‘genetically identical’ strains belonged to the same serotype, whereas genetic differences were detected between strains assigned to the same SmaI PG but differing in serotype. We conclude that PFGE-based genetic fingerprinting can yield invaluable data for epidemiological studies of sporadic C. jejuni infection, but that results based on one restriction site polymorphism must be checked with another enzyme.

Listeria monocytogenes in Five Sardinian Swine Slaughterhouses: Prevalence, Serotype, and Genotype Characterization

2013 ◽  
Vol 76 (11) ◽  
pp. 1863-1867 ◽  
Author(s):  
DOMENICO MELONI ◽  
FRANCESCA PIRAS ◽  
ANNA MUREDDU ◽  
FEDERICA FOIS ◽  
SIMONETTA GIANNA CONSOLATI ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Restriction Analysis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pork Meat ◽  
Pulsed Field ◽  
Restriction Profiles ◽  
Full Complement ◽  
Contact Surfaces ◽  
Slaughtered Pigs

In a 3-year study (2008 to 2011) to estimate the prevalence and the contamination sources of Listeria monocytogenes in pork meat in Sardinia, Italy, 211 samples were collected from five Sardinian swine slaughterhouses: 171 samples from slaughtered pigs and 40 from the slaughterhouse environment. Fifty L. monocytogenes isolates were characterized by PCR-based serotyping, presence of virulence-associated genes, and pulsed-field gel electrophoresis restriction analysis. The overall prevalence of L. monocytogenes was 33% in swine carcasses, 7% in cecal material, 23% on meat contact surfaces, and 25% on noncontact surfaces. Only two serotypes were detected: 1/2c (78%) and 1/2a (22%). In all, based on the presence of virulence-associated genes, eight pathogenic profiles were detected. Only 42% of all isolates carried the full complement of virulence-associated genes and were allotted to profile 1. Six pulsed-field gel electrophoresis profiles persisted in the slaughterhouses; restriction profiles appeared to be specific to each plant.

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