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 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 Use of Pulsed-Field Gel Electrophoresis to Determine the Source of Methicillin-Resistant Staphylococcus aureus Bacteremia

2021 ◽  
Vol 13 (3) ◽  
pp. 602-610
Author(s):  
Eugene Y. H. Yeung ◽  
Ivan Gorn
Keyword(s):  
Staphylococcus Aureus ◽  
Gel Electrophoresis ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Epidemiological Studies ◽  
Pelvic Trauma ◽  
Pulsed Field Gel Electrophoresis ◽  
Bacterial Strains ◽  
Methicillin Resistant ◽  
Pulsed Field ◽  
Clinical Cases

Pulsed-field gel electrophoresis (PFGE) has historically been considered the gold standard in fingerprinting bacterial strains in epidemiological studies and outbreak investigations; little is known regarding its use in individual clinical cases. The current study detailed two clinical cases in which PFGE helped to determine the source of their methicillin-resistant Staphylococcus aureus (MRSA) bacteremia. Patient A was found to have MRSA bacteremia after trauma in her pelvic area. MRSA was also found in her groin but not in her nostril and rectum. PFGE was performed that showed variable bands of her MRSA isolates from blood and groin, suggestive of different strains of MRSA. Her MRSA bacteremia was determined to be unrelated to her pelvic trauma. Patient B was found to have MRSA bacteremia after colonoscopy. MRSA was also found in his nostril and rectum. PFGE was performed that showed variable bands of his MRSA isolates from blood and rectum but identical bands of MRSA isolates from his blood and nostril. His MRSA bacteremia was determined to be unrelated to his colonoscopy procedure. The current study demonstrates the use of PFGE to rule out the source of bacteremia in individual clinical cases.

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 Molecular epidemiology and antimicrobial resistance of Salmonella isolates from broilers and pigs in Thailand

2019 ◽  
Vol 12 (8) ◽  
pp. 1311-1318 ◽  
Author(s):  
Dusadee Phongaran ◽  
Seri Khang-Air ◽  
Sunpetch Angkititrakul
Keyword(s):  
Antimicrobial Resistance ◽  
Gel Electrophoresis ◽  
Animal Species ◽  
Antimicrobial Agents ◽  
Genetic Relatedness ◽  
Pulsed Field Gel Electrophoresis ◽  
Resistance Pattern ◽  
Salmonella Spp ◽  
Pulsed Field ◽  
Amoxicillin Clavulanate

Aim: This study aimed to determine the prevalence and antimicrobial resistance pattern of Salmonella spp., and the genetic relatedness between isolates from broilers and pigs at slaughterhouses in Thailand. Materials and Methods: Fecal samples (604 broilers and 562 pigs) were collected from slaughterhouses from April to July 2018. Salmonella spp. were isolated and identified according to the ISO 6579:2002. Salmonella-positive isolates were identified using serotyping and challenged with nine antimicrobial agents: Amoxicillin/clavulanate (AMC, 30 μg), ampicillin (AMP, 10 μg), ceftazidime (30 μg), chloramphenicol (30 μg), ciprofloxacin (CIP, 5 μg), nalidixic acid (NAL, 30 μg), norfloxacin (10 μg), trimethoprim/sulfamethoxazole (SXT, 25 μg), and tetracycline (TET, 30 μg). Isolates of the predominant serovar Salmonella Typhimurium were examined for genetic relatedness using pulsed-field gel electrophoresis (PFGE). Results: Salmonella was detected in 18.05% of broiler isolates and 37.54% of pig isolates. The most common serovars were Kentucky, Give, and Typhimurium in broilers and Rissen, Typhimurium, and Weltevreden in pigs. Among broilers, isolates were most commonly resistant to antibiotics, NAL, AMP, TET, AMC, and CIP. Pig isolates most commonly exhibited antimicrobial resistance against AMP, TET, and SXT. Based on PFGE results among 52 S. Typhimurium isolates from broilers and pigs, a high genetic relatedness between broiler and pig isolates (85% similarity) in Cluster A and C from PFGE result was identified. Conclusion: The results revealed high cross-contamination between these two animal species across various provinces in Thailand. Keywords: antimicrobial resistance, broilers, pigs, pulsed-field gel electrophoresis, Salmonella spp.

scholarly journals Genetic Relatedness of Recently Collected Spanish Respiratory Tract Streptococcus pneumoniae Isolates with Reduced Susceptibility to Amoxicillin

2003 ◽  
Vol 47 (11) ◽  
pp. 3637-3639 ◽  
Author(s):  
E. Pérez-Trallero ◽  
J. M. Marimón ◽  
A. González ◽  
C. García-Rey ◽  
L. Aguilar
Keyword(s):  
Streptococcus Pneumoniae ◽  
Respiratory Tract ◽  
Gel Electrophoresis ◽  
Genetic Relatedness ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field

ABSTRACT Of Streptococcus pneumoniae isolates from 17 hospitals, 9.8% were amoxicillin nonsusceptible (MIC ≥ 4 μg/ml). The genetic relatedness of 138 isolates was studied by pulsed-field gel electrophoresis. Although 44 different clones were detected, more than 62% of these isolates were related to four clones (Spain23F-1, Spain6B-2, Spain9V-3, and Spain14-5).

scholarly journals Genomic pattern analysis of Burkholderia mallei field isolates by pulsed-field gel electrophoresis (PFGE) discriminatory typing

2021 ◽  
Author(s):  
Shojaat Dashtipour ◽  
Keyvan Tadayon ◽  
Sajjad Yazdansetad ◽  
Nader Mosavari ◽  
Rouhollah Keshavarz
Keyword(s):  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Outbreak Detection ◽  
Pfge Type ◽  
Identification System ◽  
Suitable Model ◽  
Burkholderia Mallei ◽  
Bacterial Isolates ◽  
Pulsed Field ◽  
Field Isolates

Background and Objectives: Glanders is a serious zoonotic disease caused by Burkholderia mallei. Prevention, control, and treatment strategies of glanders are prerequisites for microbial source tracking. The present study was aimed to analyze the genomic pattern of B. mallei Iranian field isolates by pulsed-field gel electrophoresis (PFGE) typing. Materials and Methods: B. mallei isolates were aerobically cultured in nutrient broth/agar supplemented with glycerol 4% for 48 h at 37°C. API 20NE identification system was used for the biochemical characterization. Genomic DNA of bacterial isolates was extracted using OIE-recommended protocol. Molecular identification of bacterial isolates was done based on amplification of BimA and IS407-flip genes. PFGE was applied to prepare the genomic pattern of B. mallei isolates. The guinea pig was used as a suitable model for studying the histopathological characterization of B. mallei. Results: In both enzymatic digestion patterns by using Af1II and VspI, we found three different clonal types; І) PFGE type of B. mallei Razi 325 strain, ІІ) PFGE type of Tiger, Kordan, and Oshnavieh strains, and ІІІ) PFGE type of Semirom strain. B. mallei Razi 325 was categorized as unrelated strain which was belonged to the different cluster differing more than four bands. Conclusion: PFGE showed more discriminatory power and considerable reproducibility for molecular typing of B. mallei strains in our study. It is standardized the approaches for outbreak detection, pathogen phylogeny, molecular epidemiology, and population studies.

scholarly journals Dissemination of a Chloramphenicol- and Tetracycline-Resistant but Penicillin-Susceptible Invasive Clone of Serotype 5 Streptococcus pneumoniae in Colombia

1999 ◽  
Vol 37 (7) ◽  
pp. 2337-2342 ◽  
Author(s):  
Mónica Tamayo ◽  
Raquel Sá-Leão ◽  
Ilda Santos Sanches ◽  
Elizabeth Castañeda ◽  
Hermínia de Lencastre
Keyword(s):  
Streptococcus Pneumoniae ◽  
Gel Electrophoresis ◽  
Genetic Relatedness ◽  
Invasive Disease ◽  
Pulsed Field Gel Electrophoresis ◽  
Dna Probes ◽  
National Surveillance ◽  
Surveillance Period ◽  
Pulsed Field ◽  
Serotype 5

A national surveillance conducted in Colombia between 1994 and 1996 identified serotype 5 Streptococcus pneumoniae as the second most frequent cause of invasive disease in children younger than 5 years of age. All 43 serotype 5 isolates collected during this period were shown to be susceptible to penicillin, erythromycin, cefotaxime, and vancomycin, but most (38 of 43, or 88%) were highly resistant to chloramphenicol. In order to clarify a possible genetic relatedness among these isolates, additional microbiological and molecular characterizations were performed. Most (40 of 43, or 93%) of the isolates were found to be resistant to tetracycline. Pulsed-field gel electrophoresis (PFGE) patterns of chromosomal DNAs revealed that all the 43 isolates were closely related and that 38 of the 43 isolates were representatives of a “Colombian clone” of S. pneumoniae isolates which were recovered throughout the 3-year surveillance period from patients in 13 hospitals located in five Colombian cities. Isolates belonging to this Colombian clone were resistant to chloramphenicol and tetracycline, hybridized with thecat and tetM DNA probes in the same 340-kbSmaI fragment, and had identical PFGE patterns after bothSmaI and ApaI digestions.

Sign in / Sign up

Export Citation Format

Share Document