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A Sample Extraction Method for Faster, More Sensitive PCR-Based Detection of Pathogens in Blood Culture  John F. Regan, Manohar R. Furtado, Maxim G. Brevnov,

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Presentation on theme: "A Sample Extraction Method for Faster, More Sensitive PCR-Based Detection of Pathogens in Blood Culture  John F. Regan, Manohar R. Furtado, Maxim G. Brevnov,"— Presentation transcript:

1 A Sample Extraction Method for Faster, More Sensitive PCR-Based Detection of Pathogens in Blood Culture  John F. Regan, Manohar R. Furtado, Maxim G. Brevnov, Jeanne A. Jordan  The Journal of Molecular Diagnostics  Volume 14, Issue 2, Pages (March 2012) DOI: /j.jmoldx Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

2 Figure 1 Detecting RNase P-specific target by PCR. Comparison of phenol-chloroform (A), spin column (B), and Automate Express (C) methods for purifying nucleic acids from blood samples spiked with different quantities of SPS and the use of this material in TaqMan assays for the detection of human RNase P. Samples were processed in triplicate. Data are expressed as means ± SD. IPC, internal positive control; SPS, sodium polyanetholsulfonate. The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx ) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

3 Figure 2 Detecting S. aureus-specific target by PCR. Comparison of phenol-chloroform (A), spin column (B), and Automate Express (C) methods for purifying nucleic acids from blood culture samples spiked with 10-fold dilution series of S. aureus and the use of this material in TaqMan assays for the detection of S. aureus. Each concentration of S. aureus was tested in triplicate. The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx ) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

4 Figure 3 Detecting S. aureus-specific target by PCR. Comparison of phenol-chloroform (A), spin column (B), and Automate Express (C) methods for purifying nucleic acids from different volumes of blood culture fluid samples spiked with low-titer S. aureus and the use of this material in TaqMan assays for the detection of S. aureus. Each volume was tested in triplicate. Note that the S. aureus genome weighs approximately 3 fg. The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx ) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

5 Figure 4 Detecting S. aureus-specific target by PCR. Comparison of phenol-chloroform (A), spin column (B), and Automate Express (C) methods for purifying nucleic acids from samples taken every hour from a blood culture spiked to 2060 CFU of S. aureus per milliliter and the use of this material in TaqMan assays for the detection of S. aureus. Each time point was tested in triplicate. The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx ) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

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