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 Assessment of Resolution and Intercenter Reproducibility of Results of Genotyping Staphylococcus aureus by Pulsed-Field Gel Electrophoresis of SmaI Macrorestriction Fragments: a Multicenter Study

1998 ◽  
Vol 36 (6) ◽  
pp. 1653-1659 ◽  
Author(s):  
Alex van Belkum ◽  
Willem van Leeuwen ◽  
Mary Elizabeth Kaufmann ◽  
Barry Cookson ◽  
Françoise Forey ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pfge Type ◽  
Data Sets ◽  
Bacterial Typing ◽  
Technical Problems ◽  
Pulsed Field ◽  
Pattern Similarity ◽  
Percent Similarity

Twenty well-characterized isolates of methicillin-resistantStaphylococcus aureus were used to study the optimal resolution and interlaboratory reproducibility of pulsed-field gel electrophoresis (PFGE) of DNA macrorestriction fragments. Five identical isolates (one PFGE type), 5 isolates that produced related PFGE subtypes, and 10 isolates with unique PFGE patterns were analyzed blindly in 12 different laboratories by in-house protocols. In several laboratories a standardized PFGE protocol with a commercial kit was applied successfully as well. Eight of the centers correctly identified the genetic homogeneity of the identical isolates by both the in-house and standard protocols. Four of 12 laboratories failed to produce interpretable data by the standardized protocol, due to technical problems (primarily plug preparation). With the five related isolates, five of eight participants identified the same subtype interrelationships with both in-house and standard protocols. However, two participants identified multiple strain types in this group or classified some of the isolates as unrelated isolates rather than as subtypes. The remaining laboratory failed to distinguish differences between some of the related isolates by utilizing both the in-house and standardized protocols. There were large differences in the relative genome lengths of the isolates as calculated on the basis of the gel pictures. By visual inspection, the numbers of restriction fragments and overall banding pattern similarity in the three groups of isolates showed interlaboratory concordance, but centralized computer analysis of data from four laboratories yielded percent similarity values of only 85% for the group of identical isolates. The differences between the data sets obtained with in-house and standardized protocols could be the experimental parameters which differed with respect to the brand of equipment used, imaging software, running time (20 to 48 h), and pulsing conditions. In conclusion, it appears that the standardization of PFGE depends on controlling a variety of experimental intricacies, as is the case with other bacterial typing procedures.

Human isolates ofListeria monocytogenesin Sweden during half a century (1958–2010)

2014 ◽  
Vol 142 (11) ◽  
pp. 2251-2260 ◽  
Author(s):  
G. LOPEZ-VALLADARES ◽  
W. THAM ◽  
V. SINGH PARIHAR ◽  
S. HELMERSSON ◽  
B. ANDERSSON ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pfge Type ◽  
Pulsed Field ◽  
Smoked Salmon ◽  
Soft Cheese ◽  
Cheese Production

SUMMARYIsolates ofListeria monocytogenes(n = 932) isolated in Sweden during 1958–2010 from human patients with invasive listeriosis were characterized by serotyping and pulsed-field gel electrophoresis (PFGE) (AscI). Of the 932 isolates, 183 different PFGE types were identified, of which 83 were each represented by only one isolate. In all, 483 serovar 1/2a isolates were distributed over 114 PFGE types; 90 serovar 1/2b isolates gave 32 PFGE types; 21 serovar 1/2c isolates gave nine PFGE types; three serovar 3b isolates gave one PFGE type; and, 335 serovar 4b isolates gave 31 PFGE types. During the 1980s in Sweden, several serovar 4b cases were associated with the consumption of European raw soft cheese. However, as cheese-production hygiene has improved, the number of 4b cases has decreased. Since 1996, serovar 1/2a has been the dominantL. monocytogenesserovar in human listeriosis in Sweden. Therefore, based on current serovars and PFGE types, an association between human cases of listeriosis and the consumption of vacuum-packed gravad and cold-smoked salmon is suggested.

scholarly journals Genetic characterization ofMycobacterium aviumisolates recovered from humans and animals in Australia

1996 ◽  
Vol 116 (1) ◽  
pp. 41-49 ◽  
Author(s):  
M. M. Feizabadi ◽  
I. D. Robertson ◽  
D. V. Cousins ◽  
D. Dawson ◽  
W. Chew ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Mycobacterium Avium ◽  
Gel Electrophoresis ◽  
Genetic Characterization ◽  
Genetic Relationships ◽  
Pulsed Field Gel Electrophoresis ◽  
Pfge Type ◽  
Enzyme Electrophoresis ◽  
Pulsed Field ◽  
Multilocus Enzyme Electrophoresis

SummaryGenetic relationships amongst 115 mainly Australian isolates ofMycobacterium aviumwere assessed using multilocus enzyme electrophoresis (MEE). The isolates were divided into 58 electrophoretic types (ETs), with a mean genetic diversity of 0·29. Isolates from humans were closely related to but distinct from those cultured from birds, whilst some porcine isolates belonged to the same ETs as certain human isolates. Pulsed field gel electrophoresis (PFGE) was used to differentiate related isolates, and those from birds and some from other animals, including pigs, were distinguished from the human isolates. The results of MEE and PFGE suggested that certain strains ofM. aviummay be transmitted between birds and pigs, but there was no clear evidence of transmission to humans. The serovar of theM. aviumisolates was not obviously related to their ET assignment or their PFGE type.

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 In Vitro and In Vivo Invasiveness of Different Pulsed-Field Gel Electrophoresis Types of Listeria monocytogenes

2002 ◽  
Vol 68 (11) ◽  
pp. 5698-5703 ◽  
Author(s):  
Charlotte Nexmann Larsen ◽  
Birgit Nørrung ◽  
Helle Mølgaard Sommer ◽  
Mogens Jakobsen
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
In Vivo Studies ◽  
Pfge Type ◽  
Pulsed Field ◽  
Lower Ability ◽  
Low Prevalence

ABSTRACT The virulence of different pulsed-field gel electrophoresis (PFGE) types of Listeria monocytogenes was examined by monitoring their ability to invade Caco-2 cells. Strains belonging to seven different PFGE types originating from both foods and humans were included. No significant differences in invasiveness were detected between strains isolated from humans and those isolated from food. Strains belonging to PFGE type 1 expressed a significantly lower ability to invade cells compared to strains belonging to other PFGE types. Although strains of PFGE type 2 also seemed to invade at a low level, this was not significant in the present study. PFGE types 1 and 2 as well as type 14 are more frequently found in food than the four other PFGE types examined and moreover have a relatively low prevalence in humans compared to their prevalence in food. Thus, the hypothesis that some PFGE types are less virulent than others is supported by this study showing that certain PFGE types of L. monocytogenes commonly found in food are less invasive than others to Caco-2 cells. In contrast to the differences in invasion, identical intracellular growth rates between the different PFGE types were observed. In vivo studies of the actual ability of the strains to invade the liver and spleen of cimetidine-treated rats following an oral dose of 109 L. monocytogenes cells were performed for isolates of PFGE types 1, 2, 5, and 15. After 2 days, equal amounts of bacteria were observed in the liver and spleen of the rats for any of the PFGE types tested.

PULSED-FIELD GEL ELECTROPHORESIS AS A DISCRIMINATORY TYPING TECHNIQUE FOR THE BIOTHREAT AGENT BURKHOLDERIA MALLEI

2006 ◽  
Vol 74 (3) ◽  
pp. 345-347 ◽  
Author(s):  
NARISARA CHANTRATITA ◽  
VANAPORN WUTHIEKANUN ◽  
RACHANEEPORN TIYAWISUTSRI ◽  
MONGKOL VESARATCHAVEST ◽  
TSEDEV ULZIITOGTOKH ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Burkholderia Mallei ◽  
Pulsed Field ◽  
Typing Technique
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