Current and emerging assays for Francisella tularensis detection: a review

Pohanka, Miroslav; Hubálek, Martin; Neubauerová, Vĕra; Macela, Aleš; Faldyna, Martin; Banďouchová, Hana; Pikula, Jiří

doi:10.17221/1862-vetmed

Cited by 12 publications

(9 citation statements)

References 84 publications

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“…Tularemia is considered a re-emerging zoonosis [ 1 - 3 ] that is endemic under favourable environmental conditions [ 4 ]. The highly infectious Gram-negative bacterium Francisella tularensis has been reported to cause infection in a wide range of hosts including humans [ 5 , 6 ].…”

Section: Introductionmentioning

confidence: 99%

Biochemical responses and oxidative stress in Francisella tularensis infection: a European brown hare model

et al. 2011

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BackgroundThe aim of the present study was to investigate biochemical and oxidative stress responses to experimental F. tularensis infection in European brown hares, an important source of human tularemia infections.MethodsFor these purposes we compared the development of an array of biochemical parameters measured in blood plasma using standard procedures of dry chemistry as well as electrochemical devices following a subcutaneous infection with a wild Francisella tularensis subsp. holarctica strain (a single dose of 2.6 × 109 CFU pro toto).ResultsSubcutaneous inoculation of a single dose with 2.6 × 109 colony forming units of a wild F. tularensis strain pro toto resulted in the death of two out of five hares. Plasma chemistry profiles were examined on days 2 to 35 post-infection. When compared to controls, the total protein, urea, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase were increased, while albumin, glucose and amylase were decreased. Both uric and ascorbic acids and glutathione dropped on day 2 and then increased significantly on days 6 to 12 and 6 to 14 post-inoculation, respectively. There was a two-fold increase in lipid peroxidation on days 4 to 8 post-inoculation.ConclusionsContrary to all expectations, the present study demonstrates that the European brown hare shows relatively low susceptibility to tularemia. Therefore, the circumstances of tularemia in hares under natural conditions should be further studied.

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Section: Introductionmentioning

confidence: 99%

Biochemical responses and oxidative stress in Francisella tularensis infection: a European brown hare model

et al. 2011

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“…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). cells in the sensitive area of a suitable transducer followed by formation of an immunocomplex (Fig.…”

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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“…The use of monoclonal can sometimes give poor performance compared to the polyclonal antibody. For example, a monoclonal preparation for the detection of the pathogen Francisella tularensis in the indirect format showed a difference in absorbance of 0.15 between the lowest and highest determination range whilst the polyclonal preparation gave a difference in absorbance of 0.45, with the limit of detection of 5.4×10 6 CFU/mL for polyclonal antibodies and 6.9×10 6 CFU/mL for monoclonal antibodies (Pohanka et al, 2008). The difference may be attributed to the ability of C. jejuni to exist in two different morphologies; vibrioid or bacillary forms and coccoid form, which varies according to age and stress conditions (Kelly et al, 2001).…”

Section: Detection Of C Jejuni Via Indirect Assaymentioning

confidence: 99%

The development of indirect and sandwich ELISA-based detection methods for the detection of Campylobacter jejuni using monoclonal and polyclonal antibody preparations

Masdor¹,

Altıntaş²,

Tothill³

2019

MJM

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Aims: The current gold standard method for the detection of Campylobacter jejuni is the culturing method followed up by immuno-based detection method, of which, the ELISA is the most often used. Many commercial detection methods based on ELISA use monoclonal antibody preparations although polyclonal antibody can be more sensitive and cheaper to produce. In this study, a comparison of indirect and sandwich ELISA-based detection methods for the detection of C. jejuni using a commercial monoclonal and polyclonal antibody preparations was explored. Methodology and results: An indirect and sandwich ELISA-based methods for the detection of C. jejuni was carried out using the same concentration of antibody (5 µg/mL) and the same concentration of the bacterium at 1×10 9 CFU/mL. At the pre-screening for optimum concentration of antibody to be used for both assay formats, the commercial monoclonal preparation gave a poor absorbance value of about 0.112 compared to 1.582 for the polyclonal antibody preparation. Hence, the use of the monoclonal antibody was not pursued further. Using the polyclonal antibody, the calculated Limits of Detection (LOD) value obtained for the indirect and sandwich ELISA methods were at 1.6×10 4 CFU/mL and at 1.29×10 4 CFU/mL, respectively, which are more sensitive than commercially used methods. The results of the specificity test obtained from the developed polyclonal antibody were then tested against other common food borne bacterial pathogens such as Salmonella Typhimurium, Listeria monocytogenes and Escherichia coli tested using the sandwich ELISA format indicated that the responses by other bacterial genus were relatively low with the translated cross-reactivity percentages of 1.78, 2.36, and 6.87%, respectively. Conclusion, significance and impact of study: The results indicated that the developed system using a polyclonal antibody preparation can be more sensitive than monoclonal preparation. In addition, it is also specific towards Campylobacter while the monoclonal antibody preparation fares poorly.

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Current and emerging assays for Francisella tularensis detection: a review

Cited by 12 publications

References 84 publications

Biochemical responses and oxidative stress in Francisella tularensis infection: a European brown hare model

Biochemical responses and oxidative stress in Francisella tularensis infection: a European brown hare model

Biosensors for Detection of Francisella Tularensis and Diagnosis of Tularemia

The development of indirect and sandwich ELISA-based detection methods for the detection of Campylobacter jejuni using monoclonal and polyclonal antibody preparations

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