Case Study of the Use of Pulsed Field Gel Electrophoresis in the Detection of a Food-Borne Outbreak

2015 ◽  
pp. 35-40 ◽  
Author(s):  
Niall De Lappe ◽  
Martin Cormican
Keyword(s):  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Food Borne

scholarly journals Identification and Characterization of Pathogenic Yersinia enterocolitica Isolates by PCR and Pulsed-Field Gel Electrophoresis

2005 ◽  
Vol 71 (7) ◽  
pp. 3674-3681 ◽  
Author(s):  
S. Thisted Lambertz ◽  
M.-L. Danielsson-Tham
Keyword(s):  
Multiplex Pcr ◽  
Yersinia Enterocolitica ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Virulence Plasmid ◽  
Pork Meat ◽  
Pulsed Field ◽  
Pork Products ◽  
Food Borne ◽  
Pork Samples

ABSTRACT Approximately 550 to 600 yersiniosis patients are reported annually in Sweden. Although pigs are thought to be the main reservoir of food-borne pathogenic Yersinia enterocolitica, the role of pork meat as a vehicle for transmission to humans is still unclear. Pork meat collected from refrigerators and local shops frequented by yersiniosis patients (n = 48) were examined for the presence of pathogenic Yersinia spp. A combined culture and PCR method was used for detection, and a multiplex PCR was developed and evaluated as a tool for efficient identification of pathogenic food and patient isolates. The results obtained with the multiplex PCR were compared to phenotypic test results and confirmed by pulsed-field gel electrophoresis (PFGE). In all, 118 pork products (91 raw and 27 ready-to-eat) were collected. Pathogenic Yersinia spp. were detected by PCR in 10% (9 of 91) of the raw pork samples (loin of pork, fillet of pork, pork chop, ham, and minced meat) but in none of the ready-to-eat products. Isolates of Y. enterocolitica bioserotype 4/O:3 were recovered from six of the PCR-positive raw pork samples; all harbored the virulence plasmid. All isolates were recovered from food collected in shops and, thus, none were from the patients' home. When subjected to PFGE, the six isolates displayed four different NotI profiles. The same four NotI profiles were also present among isolates recovered from the yersiniosis patients. The application of a multiplex PCR was shown to be an efficient tool for identification of pathogenic Y. enterocolitica isolates in naturally contaminated raw pork.

scholarly journals Pulsed-Field Gel Electrophoresis as a Molecular Tool for Characterizing Genomes of Certain Food-Borne Bacterial Isolates - A Review

2014 ◽  
Vol 29 ◽  
pp. 13-23 ◽  
Author(s):  
Partha Pal
Keyword(s):  
Gel Electrophoresis ◽  
Genetic Relatedness ◽  
Pulsed Field Gel Electrophoresis ◽  
Human Beings ◽  
Bacterial Isolates ◽  
Bacterial Strains ◽  
Typing Method ◽  
Gold Standard Method ◽  
Pulsed Field ◽  
Food Borne

The evolutionary transition from phenotypic to molecular analysis of infectious disease in bacterial epidemiology led to the search for suitable approaches to ascertain genomic relatedness or heterogeneity between bacterial clinical isolates. Pulsed-field gel electrophoresis (PFGE) technique was developed for separating and analyzing long DNA fragments of several megabases in alternating electric field. Comparison of electrophoresis profiles of restriction enzyme-digested genomic DNA from bacterial isolates has proved to be a useful epidemiological tool for genetic discrimination of bacterial strains, detection of genetic relatedness, to locate the source of outbreak and to monitor the spread of the microorganisms in endemic zones. PFGE is considered as a gold standard method for typing of bacterial isolates because of the remarkable endurance of this technique as a typing method for the last 20 years in molecular epidemiology. In this current review the pros and cons of PFGE use in current molecular microbiological research are explored in the context of determination of genome organization of certain food-borne bacterial isolates causing infectious diseases in human beings.

scholarly journals An outbreak of a possibly new Salmonella enterica subspecies enterica serovar with the antigenic formula 11:z41:e,n,z15, Greece, March to May 2016: preliminary results

2016 ◽  
Vol 21 (25) ◽  
Author(s):  
Georgia Mandilara ◽  
Kassiani Mellou ◽  
Kleon Karadimas ◽  
Leonidas Georgalis ◽  
Michalis Polemis ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Reference Laboratory ◽  
National Reference Laboratory ◽  
Pulsed Field ◽  
Preliminary Results ◽  
National Reference ◽  
Antigenic Type ◽  
Epidemiological Link

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.

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 Fecal Excretion of Salmonella entericaSerovar Typhimurium Following a Food-Borne Outbreak

2000 ◽  
Vol 38 (9) ◽  
pp. 3495-3497 ◽  
Author(s):  
Toshiyuki Murase ◽  
Mikiko Yamada ◽  
Tetsunori Muto ◽  
Akiyoshi Matsushima ◽  
Shiro Yamai
Keyword(s):  
Gel Electrophoresis ◽  
Salmonella Enterica ◽  
Pulsed Field Gel Electrophoresis ◽  
Fecal Excretion ◽  
Antimicrobial Drugs ◽  
Pulsed Field ◽  
Infected People ◽  
The People ◽  
Food Borne ◽  
Serovar Typhimurium

Fecal excretion of Salmonella enterica serovar Typhimurium organisms was observed in patients and in people not showing symptoms who were involved in an outbreak of food-borne infection with this organism. Excretion of organisms was prolonged in the patients who were given antimicrobial drugs compared with those who were not. The isolates were indistinguishable by their pulsed-field gel electrophoresis patterns and biotyping from the strain recovered from the roast pork that had been consumed by all of the people. This indicates that these isolates obtained from the infected people had originated in the contaminated pork.

scholarly journals Genetic relatedness of group A streptococci of the newly designated serotype M90 causing a food-borne outbreak and sporadic infections

2000 ◽  
Vol 125 (2) ◽  
pp. 299-301 ◽  
Author(s):  
S. POURNARAS ◽  
A. EFSTRATIOU ◽  
J. DOUBOYAS ◽  
R. C. GEORGE ◽  
A. TSAKRIS
Keyword(s):  
Gel Electrophoresis ◽  
Genetic Relatedness ◽  
Group A Streptococcus ◽  
Pulsed Field Gel Electrophoresis ◽  
Group A Streptococci ◽  
Pulsed Field ◽  
Food Borne ◽  
Group A ◽  
Pfge Profiles

Twenty-six isolates of the newly designated M90 serotype group A Streptococcus (GAS) from a large food-borne outbreak of pharyngitis in Greece and six M90 sporadic isolates from UK, were typed by pulsed-field gel electrophoresis (PFGE). Twenty-four outbreak isolates were identical and two closely related. The Greek isolates were possibly related with one UK isolate, while other sporadic isolates exhibited distinct PFGE profiles from the former isolates.

scholarly journals Epidemiological Study of a Food-Borne Outbreak of EnterotoxigenicEscherichia coli O25:NM by Pulsed-Field Gel Electrophoresis and Randomly Amplified Polymorphic DNA Analysis

1998 ◽  
Vol 36 (3) ◽  
pp. 652-656 ◽  
Author(s):  
Toshihiro Mitsuda ◽  
Tetsunori Muto ◽  
Mikiko Yamada ◽  
Nobuyoshi Kobayashi ◽  
Masanori Toba ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Dna Analysis ◽  
Pulsed Field Gel Electrophoresis ◽  
Causal Agent ◽  
Randomly Amplified Polymorphic Dna ◽  
Pulsed Field ◽  
Food Borne ◽  
Length Polymorphisms ◽  
Genotypic Analysis ◽  
Epidemiological Characteristics

This study investigated the applicability of molecular epidemiological techniques to the identification of the causal agent of an outbreak of diarrhea caused by ingestion of food contaminated with enterotoxigenic Escherichia coli (ETEC). The outbreak occurred at four elementary schools in July 1996 and affected more than 800 people. Illness was most strongly associated with eating tuna paste (relative risk, 1.79; 95% confidence interval = 1.16 to 2.79; P = 0.0001). To evaluate the epidemiological characteristics of the pathogen, the DNAs from numerous isolated ETEC strains were subjected to randomly amplified polymorphic DNA analysis, pulsed-field gel electrophoresis of nuclease S1-treated plasmid DNA, and analysis of genomic DNA restriction fragment length polymorphisms. All ETEC isolates were of the O25:NM (nonmotile) serotype, which carries a heat-stable enterotoxin Ib gene. Genotypic analysis demonstrated that the strains isolated from the patients at all four schools were identical. The isolates of ETEC O25:NM obtained from the tuna paste that had been served for lunch at these schools were genetically indistinguishable from those isolated from the patients. Results suggest that this outbreak was food borne. The molecular biology-based epidemiological techniques used in this study were useful in characterizing the causal agent in this food-borne epidemic.

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