Detection of Francisella tularensis in ulcers of patients with tularemia by PCR

Sjöstedt, Anders; Eriksson, Urban; Berglund, Lennart; Tärnvik, Arne

doi:10.1128/jcm.35.5.1045-1048.1997

Cited by 137 publications

(71 citation statements)

References 17 publications

(19 reference statements)

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“…After centrifugation (3000 g/10 s), the supernatant was collected and DNA was extracted with the peqGOLD TriFast Isolation System (Peqlab, Germany) following the manufacturer's instructions. The presence of B. burgdorferi s.l., A. phagocytophilum, 'Candidatus N. mikurensis', SFG rickettsiae and F. tularensis DNA in ticks was tested using PCR protocols published elsewhere (Regnery et al, 1991;Roux et al, 1996;Sj€ ostedt et al, 1997;Liebisch et al, 1998;Brown et al, 2001). Amplified PCR products were visualized by agarose gel electrophoresis, purified and directly sequenced at a commercial company (LGC Genomics, Germany).…”

Section: Methodsmentioning

confidence: 99%

First Molecular Evidence of Zoonotic Bacteria in Ticks in Bosnia and Herzegovina

Hodžić

Fuehrer

Duscher

2016

Transbound Emerg Dis

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In Bosnia and Herzegovina, the tick fauna is very diverse, but data on the occurrence of zoonotic tick-borne bacteria are lacking. Thus, the aim of this study was to investigate the presence of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, 'Candidatus Neoehrlichia mikurensis', spotted fever group (SFG) rickettsiae and Francisella tularensis in questing ticks. In 19 (21.8%) of 87 ticks (Ixodes ricinus, n = 30; Dermacentor reticulatus, n = 54; D. marginatus, n = 3) collected by flagging the vegetation at the collection site in the Glamoč Municipality (south-western Bosnia and Herzegovina), Rickettsia monacensis (1.1%), R. helvetica (5.7%), R. raoultii (5.7%), R. slovaca (8.0%), A. phagocytophilum (1.1%) and F. tularensis subsp. holartica (1.1%) were detected and identified by molecular methods. None of the tested ticks were positive for B. burgdorferi s.l. and 'Candidatus N. mikurensis', and co-infection of R. slovaca and F. tularensis subsp. holarctica was detected in only one D. marginatus (1.1%). This study reports the occurrence of emerging zoonotic bacteria in ticks from Bosnia and Herzegovina for the first time, indicating a public health threat to humans. Therefore, physicians and practitioners should be aware of the presence of these tick-borne bacteria, especially when they are faced with acute febrile illnesses after tick exposure.

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Section: Methodsmentioning

confidence: 99%

First Molecular Evidence of Zoonotic Bacteria in Ticks in Bosnia and Herzegovina

Hodžić

Fuehrer

Duscher

2016

Transbound Emerg Dis

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“…However, limited clinical diagnostic data for the Francisella ELISA do not allow for assessment of the diagnostic sensitivity and specificity of the Francisella ABICAP system based on this assay or other approaches such as PCR or culturing on appropriate growth media as a reference. In one study, a diagnostic sensitivity of PCR of approximately 73% for patients with ulceroglandular tularemia has been shown (Sjostedt et al 1997).…”

mentioning

confidence: 99%

RAPID DETECTION OF FRANCISELLA TULARENSIS BY THE IMMUNOAFFINITY ASSAY ABICAP IN ENVIRONMENTAL AND HUMAN SAMPLES

Grunow

Miethe²,

Conlan

et al. 2008

Rapid Methods Automation Mic

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Francisella tularensis is the causative agent of tularemia, a severe zoonotic infectious disease. The natural reservoir of the bacteria is not yet known precisely, but it may persist for over a year in water or mud. It is naturally maintained and spread by various terrestrial and aquatic mammals, and outbreaks in humans are often paralleled by enzootics or epizootics. The reasons for this seem to be multifactorial, though not well understood. Therefore, for epidemiological and outbreak investigations, a rapid hand‐held assay would be helpful. Here we described a column‐based immunofiltration assay called ABICAP, which has several characteristics that permit its application under field conditions. The assay can be performed within 25 min, with a detection limit of 103 bacteria. Further validation of the assay included the analytical range, precision, detection limit and performance for different human, animal and environmental sample matrices, accuracy, recovery, reproducibility and stability. PRACTICAL APPLICATIONS The assay is an interesting tool applicable for the rapid detection of F. tularensis in various specimens under laboratory and field conditions. It can be used for the identification of reservoirs of F. tularensis in epidemiological studies, for the detection of sources of infection in outbreak situations of tularemia and in laboratory investigation of animal and human specimens. The methodological platform can be adapted to the detection of other microbiological agents and antigens.

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“…Different methods are available for detecting microbial agents. Some of these methods are culturedependent with infection risks, whereas some of them are time-consuming immunoassays (19,20), requiring advanced equipment. Correct identification of these pathogens depends on culturing and genome extraction, for which level 3 laboratories and specialized researchers are necessary.…”

Section: Discussionmentioning

confidence: 99%

Simple and Rapid Detection of Yersinia pestis and Francisella tularensis Using Multiplex-PCR: Molecular Detection of Yersinia pestis and Francisella tularensis

Pourmahdi

Zeinoddini

Esmatabadi

et al. 2019

RMM

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Background: Yersinia pestis and Francisella tularensis cause plague and tularemia, which are known as diseases of the newborn and elderly, respectively. Immunological and culture-based detection methods of these bacteria are time-consuming, costly, complicated and require advanced equipment. We aimed to design and synthesize a gene structure as positive control for molecular detection of these bacteria. Materials and Method: Conserved regions of each bacterium were determined. A fragment containing the fopA and caf1 genes (conserved genes of F. tularensis and Y. pestis, respectively) was artificially synthesized, cloned into the pUC57 vector (pUC-fopA-caf1), transformed into E. coli DH5α, and used in a multiplex PCR assay. The sensitivity of this assay was examined by serial dilution of the extracted plasmid, whereas the specificity was examined using genomes of Escherichia coli, Salmonella typhi, Enterobacter aerogenes, Vibrio cholerae as templates. Finally, PCR products were analyzed in agarose gel electrophoresis. Results: As expected, our analysis showed a clear dual band in the size range of 107 bp to 176 bp, confirming the presence of fopA and caf1 genes. Another 351 bp band was detected due to amplification being dependent on the forward primer of fopA and the reverse primer of caf1. Optimization of the PCR protocol reduced the amplification of this 351 bp band. The sensitivity of this assay was determined to be 36×10 −3 ng/µl and the selectivity test confirmed the specificity of this method is appropriate for the detection of target genes. Conclusion: This multiplex PCR method could be used in research laboratories for identification of these important pathogens.

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Detection of Francisella tularensis in ulcers of patients with tularemia by PCR

Cited by 137 publications

References 17 publications

First Molecular Evidence of Zoonotic Bacteria in Ticks in Bosnia and Herzegovina

First Molecular Evidence of Zoonotic Bacteria in Ticks in Bosnia and Herzegovina

RAPID DETECTION OF FRANCISELLA TULARENSIS BY THE IMMUNOAFFINITY ASSAY ABICAP IN ENVIRONMENTAL AND HUMAN SAMPLES

Simple and Rapid Detection of Yersinia pestis and Francisella tularensis Using Multiplex-PCR: Molecular Detection of Yersinia pestis and Francisella tularensis

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