Using real-time PCR to specifically detect Burkholderia mallei

Ulrich, Melanie; Norwood, David; Christensen, Deanna R.; Ulrich, Ricky L.

doi:10.1099/jmm.0.46350-0

Cited by 35 publications

(25 citation statements)

References 29 publications

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“…The real-time PCR assays used in this study have been used by USAMRIID for the in vitro qualitative presumptive detection of BSAT DNA in environmental samples, clinical specimens and culture isolates and have been described in previous reports [13][14][15]. We used the following four primer and probe sets: BALB (chrom.…”

Section: Real-time Pcrmentioning

confidence: 99%

“…3.3)]. , using the real-time PCR assays described by Bode et al [13], Christensen et al [14] and Ulrich et al [15] with the LightCycler FastStart DNA Master HybProbe (Roche I). Serial dilutions of purified bacterial DNA from each bioterrorism threat agent were prepared, and the assays were performed in triplicate with samples at each concentration on the 7500 Fast Dx instrument.…”

Section: Comparison Of the Ten Master Mixes On The 7500 Fast DX Instrmentioning

confidence: 99%

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Multi-platform comparison of ten commercial master mixes for probe-based real-time polymerase chain reaction detection of bioterrorism threat agents for surge preparedness

Buzard

Baker

Wolcott

et al. 2012

Forensic Science International

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Section: Real-time Pcrmentioning

confidence: 99%

Section: Comparison Of the Ten Master Mixes On The 7500 Fast DX Instrmentioning

confidence: 99%

Multi-platform comparison of ten commercial master mixes for probe-based real-time polymerase chain reaction detection of bioterrorism threat agents for surge preparedness

Buzard

Baker

Wolcott

et al. 2012

Forensic Science International

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“…DNA from the swab diluents, BAL wash fluid, tissue samples, whole blood and serum was extracted using a Qiagen DNA blood kit according to the manufacturer's instructions. PCR was performed on a LightCycler 1.5 Real-Time PCR instrument (Roche) with an assay specific for B. mallei, as well as a more sensitive assay that detected both B. mallei and B. pseudomallei (IS407A insertion element) (Ulrich et al, 2006). Assays were carried out in 20 ml volumes.…”

Section: Methodsmentioning

confidence: 99%

“…Each assay contained 1|PCR buffer (50 mM Tris, pH 8.3, 250 mg BSA ml 2 1 ) (Idaho Technology), 0.2 mM dNTP mix, 1.0 U Platinum Taq DNA polymerase (Invitrogen) and 5 mM MgCl 2 . Previously determined optimal concentrations of primers and probe were added, and 15 ml master mix was distributed to reaction tubes (Ulrich et al, 2006). An aliquot of 5 ml control/template/sample DNA was added just before analysis on the instrument.…”

Section: Methodsmentioning

confidence: 99%

Pathological findings and diagnostic implications of a rhesus macaque (Macacca mulatta) model of aerosol exposure to Burkholderia mallei (glanders)

Yingst

Facemire

Chuvala

et al. 2015

Journal of Medical Microbiology

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Burkholderia mallei is a Gram-negative bacillus that causes a pneumonic disease known as glanders in equids and humans, and a lymphatic infection known as farcy, primarily in equids. With the potential to infect humans by the respiratory route, aerosol exposure can result in severe, occasionally fatal, pneumonia. Today, glanders infections in humans are rare, likely due to less frequent contact with infected equids than in the past. Acutely ill humans often have non-specific clinical signs and in order to diagnose cases, especially in scenarios of multiple cases in an unexpected setting, rapid diagnostics for B. mallei may be critical. The pathogenesis of acute glanders in the rhesus macaque (Macaca mulatta) was studied as an initial effort to improve diagnostic methods. In the study described here, the diagnostic techniques of PCR, culture and histopathology were compared. The results indicated that PCR may provide rapid, non-invasive diagnosis of glanders in some cases. As expected, PCR results were positive in lung tissue in 11/12 acutely infected rhesus macaques, but more importantly in terms of diagnostic algorithm development, PCR results were frequently positive in non-invasive samples such as broncho-alveolar lavage or nasal swabs (7/12) and occasionally in blood (3/12). However, conventional bacterial culture failed to recover bacteria in many of these samples. The study showed that the clinical presentation of aerosol-exposed rhesus macaques is similar to descriptions of human glanders and that PCR has potential for rapid diagnosis of outbreaks, if not individual cases.

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“…Despite the advantages of both PCR and bacterial culture, each technology still requires a circulating agent for detection/identification to be facilitated. Some agents that cause acute disease, such as Burkholderia mallei (Ulrich et al, 2006), do not result in high bacterial blood titres in infected individuals. Therefore, an appropriate, uniform, clinical sample type for all PCRs on a multiplexed platform consumable may not be available.…”

Section: S a Weller And Othersmentioning

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

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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).

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Using real-time PCR to specifically detect Burkholderia mallei

Cited by 35 publications

References 29 publications

Multi-platform comparison of ten commercial master mixes for probe-based real-time polymerase chain reaction detection of bioterrorism threat agents for surge preparedness

Multi-platform comparison of ten commercial master mixes for probe-based real-time polymerase chain reaction detection of bioterrorism threat agents for surge preparedness

Pathological findings and diagnostic implications of a rhesus macaque (Macacca mulatta) model of aerosol exposure to Burkholderia mallei (glanders)

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

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