Development of real-time PCR assays for the specific detection of Francisella tularensis ssp. tularensis, holarctica and mediaasiatica

Mitchell, Jan L.; Chatwell, Nicola; Christensen, Deanna R.; Diaper, Helen; Minogue, Timothy D.; Parsons, Tanya M.; Walker, Brian; Weller, Simon A.

doi:10.1016/j.mcp.2009.10.004

Cited by 24 publications

(20 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…CR is used to detect biological warfare agents (BWAs) from various sample types (1)(2)(3)(4)(5). In this context, multiple PCR inhibitors negatively affect agent detection; these are known to include compounds such as humic acids, hemoglobin, complex polysaccharides, hematin, and urea (6)(7)(8)(9)(10)(11).…”

mentioning

confidence: 99%

Cross-Institute Evaluations of Inhibitor-Resistant PCR Reagents for Direct Testing of Aerosol and Blood Samples Containing Biological Warfare Agent DNA

Minogue

Rachwal

Hall

et al. 2014

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

The down-selected reagents underwent further testing. In the United Kingdom experiments, both reagents were tested against seven contrived aerosol collector samples containing B. anthracis Ames DNA and B. subtilis spores from a commercial formulation (BioBall). In PCR assays with reaction mixtures containing 40% crude sample, an airfield-collected sample induced inhibition of the B. subtilis PCR with the KAPA reagent and complete failure of both PCRs with the Fast Virus reagent. However, both reagents allowed successful PCR for all other samples-which inhibited PCRs with a non-inhibitor-resistant reagent. In the United States, a cross-assay limit-of-detection (LoD) study in blood was conducted. The KAPA Blood Direct reagent allowed the detection of agent DNA (by four PCRs) at higher concentrations of blood in the reaction mixture (2.5%) than the Fast Virus reagent (0.5%), although LoDs differed between assays and reagent combinations. Across both groups, the KAPA Blood Direct reagent was determined to be the optimal reagent for inhibition relief in PCR.

show abstract

mentioning

confidence: 99%

Cross-Institute Evaluations of Inhibitor-Resistant PCR Reagents for Direct Testing of Aerosol and Blood Samples Containing Biological Warfare Agent DNA

Minogue

Rachwal

Hall

et al. 2014

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

show abstract

“…The development and validation of the F. tularensis PCRs have been described previously (Mitchell et al, 2010), and the performance of all seven PCRs in singleplex and Array Card format has also been reported (Rachwal et al, 2012). All B. anthracis and Y. pestis PCR assays have been shown to be specific for their target genes/agents with the exception of the Y. pestis mpbA assay, which has been shown also to detect Yersinia pseudotuberculosis.…”

Section: Methodsmentioning

confidence: 99%

“…PCR is commonly used to detect many bacterial pathogens from different sample types (Mitchell et al, 2010;Rose et al, 2011;Weller et al, 2000). A PCR assay asks a biased question of a sample, i.e.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of two multiplex real-time PCR screening capabilities for the detection of Bacillus anthracis, Francisella tularensis and Yersinia pestis in blood samples generated from murine infection models

Weller

Cox

Essex-Lopresti

et al. 2012

Journal of Medical Microbiology

Self Cite

View full text Add to dashboard Cite

Two multiplex PCR screening capabilities (TaqMan Array Cards and FilmArray) were evaluated for their ability to detect Bacillus anthracis, Francisella tularensis and Yersinia pestis in blood samples obtained from respective murine infection models. Blood samples were obtained from infected mice at 24 h intervals after exposure. Multiplex PCR results were compared with standard blood culture and singleplex real-time PCR. Across all three models, 71 mice were tested in total, within which a subset of 43 samples was shown to contain an infecting agent by at least one of the detection technologies. Within this subset of positive samples, for each model studied, the detection rates of each technology were compared. The B. anthracis model blood culture (14 of 15 agent-containing samples tested) and FilmArray PCR (12 of 15) were shown to have equivalent detection rates, which were significantly higher (at the 95 % confidence level) than singleplex (five of 14) or Array Card (two of 14) PCRs. The F. tularensis model blood culture (12 of 12) was shown to have a significantly higher (at 95 % confidence level) detection rate than all PCR technologies, with FilmArray (seven of 11) and singleplex (seven of 12) PCRs shown to have significantly higher (at 95 % confidence level) detection rates than the Array Card PCR (two of 11). Within the Y. pestis model, there was no significant difference in detection rates between blood culture (10 of 16), singleplex PCR (14 of 16), Array Card PCR (10 of 16) and FilmArray PCR (10 of 13).

show abstract

“…The Polymerase Chain Reaction (PCR) is commonly used to detect many bacterial pathogens from different sample types [6,7] and can achieve a result within 2 hours. Detection of B. anthracis is often facilitated by PCR assays designed against genetic targets on the virulence associated pXO1 and pXO2 plasmids.…”

Section: Introductionmentioning

confidence: 99%

Development of Three Real-Time PCR assays to Detect Bacillus anthracis and Assessment of Diagnostic Utility

Parsons¹

2013

J Bioterr Biodef

View full text Add to dashboard Cite

Development of real-time PCR assays for the specific detection of Francisella tularensis ssp. tularensis, holarctica and mediaasiatica

Cited by 24 publications

References 21 publications

Cross-Institute Evaluations of Inhibitor-Resistant PCR Reagents for Direct Testing of Aerosol and Blood Samples Containing Biological Warfare Agent DNA

Cross-Institute Evaluations of Inhibitor-Resistant PCR Reagents for Direct Testing of Aerosol and Blood Samples Containing Biological Warfare Agent DNA

Evaluation of two multiplex real-time PCR screening capabilities for the detection of Bacillus anthracis, Francisella tularensis and Yersinia pestis in blood samples generated from murine infection models

Development of Three Real-Time PCR assays to Detect Bacillus anthracis and Assessment of Diagnostic Utility

Contact Info

Product

Resources

About