Recovery of Francisella tularensis from soil samples by filtration and detection by real-time PCR and cELISA

Jiménez, Óscar Esteban; Aizpurua, Carmen; Fernandez-Frutos, Begoña; León, Patricia de; Camacho, Maite; Fernández-Moreira, Daniel; Ybarra, Carmen; Cabria, Juan Carlos

doi:10.1039/b716608g

Cited by 11 publications

(9 citation statements)

References 34 publications

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“…tularensis (Ftt) was developed as a potential biological weapon, and has since been categorized as a Tier 1 select agent in the USA [4]. Ftt is readily found in the environment as both a free-living and arthropodtransmitted pathogen [5,6]. Thus, this bacterium also represents a persistent public health threat in both developed and underdeveloped areas of the world [7,8].…”

Section: Introductionmentioning

confidence: 99%

Unique Francisella Phosphatidylethanolamine Acts as a Potent Anti-Inflammatory Lipid

Ireland¹,

Schwarz²,

Nardone³

et al. 2018

J Innate Immun

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Virulent Francisella tularensis subsp. tularensis (Ftt) is a dynamic, intracellular, bacterial pathogen. Its ability to evade and rapidly suppress host inflammatory responses is considered a key element for its profound virulence. We previously established that Ftt lipids play a role in inhibiting inflammation, but we did not determine the lipid species mediating this process. Here, we show that a unique, abundant, phosphatidylethanolamine (PE), present in Francisella, contributes to driving the suppression of inflammatory responses in human and mouse cells. Acyl chain lengths of this PE, C24: 0 and C10: 0, were key to the suppressive capabilities of Francisella PE. Addition of synthetic PE 24: 0–10: 0 resulted in the accumulation of PE in host cells for up to 24 h of incubation, and recapitulated the inhibition of inflammatory responses observed with native Ftt PE. Importantly, this novel PE significantly inhibited inflammatory responses driven by a medically and globally important flavivirus, dengue fever virus. Thus, targeting these lipids and/or the pathways that they manipulate represents a new strategy to combat immunosuppression engendered by Ftt, but they also show promise as a novel therapeutic intervention for significant viral infections.

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Section: Introductionmentioning

confidence: 99%

Unique Francisella Phosphatidylethanolamine Acts as a Potent Anti-Inflammatory Lipid

Ireland¹,

Schwarz²,

Nardone³

et al. 2018

J Innate Immun

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“…On the other hand, qPCR has been used to detect and quantify various bacteria species present in different types of samples (Coelho et al, 2009;Jung et al, 2010;Lacava et al, 2006;Ruppel et al, 2006;Sellek et al, 2008;Timmusk et al, 2009;Trabelsi et al, 2009) and therefore may represent an alternative means for quantifying G. diazotrophicus.…”

Section: Discussionmentioning

confidence: 99%

“…QPCR has been applied to quantify bacteria directly in pure culture, soil samples (Sellek et al, 2008;Trabelsi et al, 2009) and plant tissues (Lacava et al, 2006;Ruppel et al, 2006;Trabelsi et al, 2009) using either 16S rRNA (Jung et al, 2010), nifH gene (Coelho et al, 2009) or species-specific sequences (Jung et al, 2010;Timmusk et al, 2009). …”

Section: Introductionmentioning

confidence: 99%

Development of a real-time PCR assay for the detection and quantification of Gluconacetobacter diazotrophicus in sugarcane grown under field conditions

Paulo¹,

Jean²,

Leona³

et al. 2014

Afr. J. Microbiol. Res.

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The objective of this study was to evaluate the viability of real-time polymerase chain reaction (PCR) to detect Gluconacetobacter diazotrophicus in sugarcane inoculated and non-inoculated with diazotrophs which are grown under field conditions. The primer pair PAL5F and PAL5R yielded a specific band of 189 bp using real time PCR with SYBER Green I. This primer pair was the most sensitive one to detect endophytic bacteria in sugarcane plants grown under field conditions and inoculated or not with bacterium. The lower limit of detection was 5 fg of template DNA, which corresponds to 12 bacterial cells. In contrast, a cultivation-dependent approach was not capable of detecting the bacteria in the same sample. The quantification of G. diazotrophicus from field grown plants using real-time PCR and a set of specific primers can be used to determine the number of bacterial cells that colonize endophytically the plant after inoculation. A highly sensitive and specific assay was developed to quantify G. diazotrophicus in sugarcane plants grown under field conditions. This assay can be used to evaluate the occurrence of the bacterium in different sample types.

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“…In the case of soil samples, the lowest limit of detection reached only 20 CFU per 1 g of soil (Whitehouse & Hottel, 2006). The real-time PCR based on SYBR Green I and tul4 gene for F. tularensis LVS achieved limit of detection of 0.69 fg of genomic DNA (Sellek et al, 2008). An extensive review of detection methods was published recently (Pohanka et al, 2008).…”

Section: Assay Methodsmentioning

confidence: 99%

Biosensors for Detection of Francisella Tularensis and Diagnosis of Tularemia

Skládal¹,

Pohanka²

2010

Biosensors

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A biosensor is defined as a detecting device that combines a transducer with a biologically sensitive and selective component. When a specific target molecule interacts with the biological component, a signal is produced, at transducer level, proportional to the concentration of the substance. Therefore biosensors can measure compounds present in the environment, chemical processes, food and human body at low cost if compared with traditional analytical techniques. Bringing together researchers from 11 different countries, this book covers a wide range of aspects and issues related to biosensor technology, such as biosensor applications in the fields of drug discovery, diagnostics and bacteria detection, optical biosensors, biotelemetry and algorithms applied to biosensing.

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Recovery of Francisella tularensis from soil samples by filtration and detection by real-time PCR and cELISA

Cited by 11 publications

References 34 publications

Unique <b><i>Francisella</i></b> Phosphatidylethanolamine Acts as a Potent Anti-Inflammatory Lipid

Unique <b><i>Francisella</i></b> Phosphatidylethanolamine Acts as a Potent Anti-Inflammatory Lipid

Development of a real-time PCR assay for the detection and quantification of Gluconacetobacter diazotrophicus in sugarcane grown under field conditions

Biosensors for Detection of Francisella Tularensis and Diagnosis of Tularemia

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