Specific detection of Plesiomonas shigelloides isolated from aquatic environments, animals and human diarrhoeal cases by PCR based on 23S rRNA gene

González-Rey, Carlos

doi:10.1016/s0928-8244(00)00194-2

Cited by 21 publications

(22 citation statements)

References 6 publications

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“…1), corresponding to the 284-bp PCR product expected (González-Rey et al, 2000). The 3 positive control strains also yielded one band of the same size.…”

Section: Confirmation Of the Plesiomonas Shigelloides Isolates By Pcrmentioning

confidence: 86%

“…The primer pair PS23FW3 and PS23RV3 are specific for P. shigelloides and delineate a 284-bp sequence of the 23S rRNA gene (González-Rey et al, 2000). Primers LMPB1 and LMPB4 (Table 2) are random primers originally described for RAPD analysis of L. monocytogenes by Boerlin et al (1995).…”

Section: Primersmentioning

confidence: 99%

“…The total PCR volume was 50 μl and (Techne Inc., Princeton, NJ, USA) with the following PCR temperature-cycling parameters: 1 cycle at 95°C for 5 min followed by 35 cycles with a denaturation step at 94°C for 1 min, an annealing temperature of 68°C for 1 min and extension at 72°C for 1 min. A final extension step at 72°C was performed at the end of the 35 cycles for 10 min (González-Rey et al, 2000). Electrophoresis of the PCR products was performed in 1.0% agarose gels (cat.…”

Section: Pcr Confirmation Of P Shigelloidesmentioning

confidence: 99%

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Genetic Variability Among Isolates ofPlesiomonas shigelloidesfrom Fish, Human Clinical Sources and Fresh Water, Determined by RAPD Typing

González-Rey²,

Krovacek³

et al. 2006

Food Biotechnology

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Plesiomonas shigelloides has been a suspected pathogen for the past 40 years. Random amplified polymorphic DNA (RAPD) analysis with two random primers was used to examine the genetic diversity among 26 strains of P. shigelloides: 10 from fresh water, 6 from fish, and 10 from human clinical sources. There was notable genetic variability among most of the isolates, and none of the isolates had the same composite RAPD profile. Our results indicate that most of the isolates from the same source grouped together and that the isolates from fish had a closer linkage to the human clinical isolates than did the freshwater isolates, suggesting that fish may be the more serious source for potential risk of infection. We also found that certain human clinical isolates had almost the same RAPD profiles as certain of the isolates from freshwater and fish, indicating that P. shigelloides from both sea food and freshwater should be considered potential pathogens.

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“…1), corresponding to the 284-bp PCR product expected (González-Rey et al, 2000). The 3 positive control strains also yielded one band of the same size.…”

Section: Confirmation Of the Plesiomonas Shigelloides Isolates By Pcrmentioning

confidence: 86%

Section: Primersmentioning

confidence: 99%

Section: Pcr Confirmation Of P Shigelloidesmentioning

confidence: 99%

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Genetic Variability Among Isolates ofPlesiomonas shigelloidesfrom Fish, Human Clinical Sources and Fresh Water, Determined by RAPD Typing

González-Rey²,

Krovacek³

et al. 2006

Food Biotechnology

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“…1) in order to maximize the diversity of our population. Isolates that were gram negative and oxidase, indole, maltose, and glucose positive were further identified by biochemical typing using the API 20 E system (bioMerieux SA, Marcy-l'Etoile, France) and were tested by specific PCR for detection of P. shigelloides based on the 23S rRNA gene as previously described (26).…”

Section: Methodsmentioning

confidence: 99%

“…P. shigelloides strains can be identified using phenotypic properties (3,34). Specific PCR amplification of the 23S rRNA and hugA genes was designed for identification purposes (26,30).…”

mentioning

confidence: 99%

Recombining Population Structure of Plesiomonas shigelloides ( Enterobacteriaceae ) Revealed by Multilocus Sequence Typing

et al. 2007

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Plesiomonas shigelloides is an emerging pathogen that is widespread in the aquatic environment and is responsible for intestinal diseases and extraintestinal infections in humans and other animals. Virtually nothing is known about its genetic diversity, population structure, and evolution, which severely limits epidemiological control. We addressed these questions by developing a multilocus sequence typing (MLST) system based on five genes (fusA, leuS, pyrG, recG, and rpoB) and analyzing 77 epidemiologically unrelated strains from several countries and several ecological sources. The phylogenetic position of P. shigelloides within family Enterobacteriaceae was precisely defined by phylogenetic analysis of the same gene portions in other family members. Within P. shigelloides, high levels of nucleotide diversity (average percentage of nucleotide differences between strains, 1.49%) and genotypic diversity (64 distinct sequence types; Simpson's index, 99.7%) were found, with no salient internal phylogenetic structure. We estimated that homologous recombination in housekeeping genes affects P. shigelloides alleles and nucleotides 7 and 77 times more frequently than mutation, respectively. These ratios are similar to those observed in the naturally transformable species Streptococcus pneumoniae with a high rate of recombination. In contrast, recombination within Salmonella enterica, Escherichia coli, and Yersinia enterocolitica was much less frequent. P. shigelloides thus stands out among members of the Enterobacteriaceae. Its high rate of recombination results in a lack of association between genomic background and O and H antigenic factors, as observed for the 51 serotypes found in our sample. Given its robustness and discriminatory power, we recommend MLST as a reference method for population biology studies and epidemiological tracking of P. shigelloides strains.Plesiomonas shigelloides is a species of rod-shaped gramnegative bacteria recently classified in the family Enterobacteriaceae and is the only oxidase-positive member of this family (32). Freshwater and estuarine water are considered to be the natural environment of P. shigelloides (36), which is often isolated from fish and seafood (30,42). Like many Enterobacteriaceae species, P. shigelloides is also found in a wide range of hosts, including dogs,

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Rapid and Specific Detection of Plesiomonas shigelloides Directly from Stool by LightCycler PCR

Loh¹,

Yap

2002

Rapid Cycle Real-Time PCR — Methods and Applications

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Specific detection of Plesiomonas shigelloides isolated from aquatic environments, animals and human diarrhoeal cases by PCR based on 23S rRNA gene

Cited by 21 publications

References 6 publications

Genetic Variability Among Isolates ofPlesiomonas shigelloidesfrom Fish, Human Clinical Sources and Fresh Water, Determined by RAPD Typing

Genetic Variability Among Isolates ofPlesiomonas shigelloidesfrom Fish, Human Clinical Sources and Fresh Water, Determined by RAPD Typing

Recombining Population Structure of Plesiomonas shigelloides ( Enterobacteriaceae ) Revealed by Multilocus Sequence Typing

Rapid and Specific Detection of Plesiomonas shigelloides Directly from Stool by LightCycler PCR

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