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,

Slides:

Advertisements

Similar presentations

Measurement of Relative Copy Number of CDKN2A/ARF and CDKN2B in Bladder Cancer by Real-Time Quantitative PCR and Multiplex Ligation-Dependent Probe Amplification.

Advertisements

CyclinD1/CyclinD3 Ratio by Real-Time PCR Improves Specificity for the Diagnosis of Mantle Cell Lymphoma Carol D. Jones, Katherine H. Darnell, Roger A.

Quantification of Parvovirus B19 DNA Using COBAS AmpliPrep Automated Sample Preparation and LightCycler Real-Time PCR Stefan Schorling, Gunnar Schalasta,

Design and Multiseries Validation of a Web-Based Gene Expression Assay for Predicting Breast Cancer Recurrence and Patient Survival Ryan K. Van Laar The.

Direct DNA Amplification from Crude Clinical Samples Using a PCR Enhancer Cocktail and Novel Mutants of Taq Zhian Zhang, Milko B. Kermekchiev, Wayne.

Multicenter Evaluation of the LightCycler MRSA Advanced Test, the Xpert MRSA Assay, and MRSASelect Directly Plated Culture with Simulated Workflow Comparison.

Carol Beadling, Tanaya L. Neff, Michael C

Direct DNA Amplification from Crude Clinical Samples Using a PCR Enhancer Cocktail and Novel Mutants of Taq Zhian Zhang, Milko B. Kermekchiev, Wayne.

Performance Characteristics of a Quantitative Hepatitis C Virus RNA Assay Using COBAS AmpliPrep Total Nucleic Acid Isolation and COBAS TaqMan Hepatitis.

Real-Time Polymerase Chain Reaction for Detecting Bacterial DNA Directly from Blood of Neonates Being Evaluated for Sepsis Jeanne A. Jordan, Mary Beth.

Comparative Study and Analytical Verification of PCR Methods for the Diagnosis of Congenital Chagas Disease Carolina I. Cura, Juan C. Ramírez, Marcelo.

Laboratory Guidelines for Detection, Interpretation, and Reporting of Maternal Cell Contamination in Prenatal Analyses Narasimhan Nagan, Nicole E. Faulkner,

A Novel Technology for Multiplex Gene Expression Analysis Directly from Whole Blood Samples Stabilized at Ambient Temperature Using an RNA-Stabilizing.

Comparison of BIOMED-2 Versus Laboratory-Developed Polymerase Chain Reaction Assays for Detecting T-Cell Receptor-γ Gene Rearrangements Keyur P. Patel,

A DNA Real-Time Quantitative PCR Method Suitable for Routine Monitoring of Low Levels of Minimal Residual Disease in Chronic Myeloid Leukemia Paul A.

Detection of Cytomegalovirus in Whole Blood Using Three Different Real-Time PCR Chemistries Erica Vincent, Zhengming Gu, Markus Morgenstern, Candace.

Laboratory Diagnosis of Clostridium difficile Infection

Joanna Wang, Chetan Bettegowda The Journal of Molecular Diagnostics

Increased Sensitivity of the Roche COBAS AMPLICOR HCV Test, Version 2

Maxim B. Freidin, Neesa Bhudia, Eric Lim, Andrew G

Comparison of High-Resolution Melting Analysis, TaqMan Allelic Discrimination Assay, and Sanger Sequencing for Clopidogrel Efficacy Genotyping in Routine.

Evaluation and Clinical Validation of Two Field–Deployable Reverse Transcription- Insulated Isothermal PCR Assays for the Detection of the Middle East.

Flexible Automated Platform for Blood Group Genotyping on DNA Microarrays Sandra Paris, Dominique Rigal, Valérie Barlet, Martine Verdier, Nicole Coudurier,

Combined Molecular Gram Typing and High-Resolution Melting Analysis for Rapid Identification of a Syndromic Panel of Bacteria Responsible for Sepsis-Associated.

Plasmodium Detection and Differentiation by Direct-on-Blood PCR Nucleic Acid Lateral Flow Immunoassay Johanna M. Roth, Laura de Bes, Patrick Sawa, George.

Cornelis J. J. Huijsmans, Jeroen Poodt, Paul H. M. Savelkoul, Mirjam H

Ken B. Waites, Li Xiao, Vanya Paralanov, Rose M. Viscardi, John I

A Lean Laboratory The Journal of Molecular Diagnostics

Patrick R. Murray The Journal of Molecular Diagnostics

William L. Gerald, M.D., Ph.D, 1954–2008

Evaluation of Nanofluidics Technology for High-Throughput SNP Genotyping in a Clinical Setting Maurice Chan, Mei Wen Chan, Ting Wei Loh, Hai Yang Law,

Real-Time Polymerase Chain Reaction for Detecting Bacterial DNA Directly from Blood of Neonates Being Evaluated for Sepsis Jeanne A. Jordan, Mary Beth.

Feasibility of a Novel Approach for Rapid Detection of Simulated Bloodstream Infections via Enzymatic Template Generation and Amplification (ETGA)–Mediated.

A Convenient Nucleic Acid Test on the Basis of the Capillary Convective PCR for the On-Site Detection of Enterovirus 71 Shiyin Zhang, Yanyan Lin, Jin.

Benjamin P. Song, Surbhi Jain, Selena Y. Lin, Quan Chen, Timothy M

Olivier Gruselle, Thierry Coche, Jamila Louahed

Comparative Evaluation of Four Real-Time PCR Methods for the Quantitative Detection of Epstein-Barr Virus from Whole Blood Specimens Daelynn Buelow,

Comparison of QIAsymphony Automated and QIAamp Manual DNA Extraction Systems for Measuring Epstein-Barr Virus DNA Load in Whole Blood Using Real-Time.

Comparative Evaluation of Broad-Panel PCR Assays for the Detection of Gastrointestinal Pathogens in Pediatric Oncology Patients Zhengming Gu, Haiqing.

The Molecular Pathology of Primary Immunodeficiencies

Modified Real-Time PCR for Detecting, Differentiating, and Quantifying Ureaplasma urealyticum and Ureaplasma parvum Ellen Vancutsem, Oriane Soetens,

Highly Sensitive Droplet Digital PCR Method for Detection of EGFR-Activating Mutations in Plasma Cell–Free DNA from Patients with Advanced Non–Small Cell.

Development and Laboratory Evaluation of a Real-Time PCR Assay for Detecting Viruses and Bacteria of Relevance for Community-Acquired Pneumonia Alicia.

Diagnosis of Trypanosomatid Infections

The Effect of Nucleic Acid Extraction Platforms and Sample Storage on the Integrity of Viral RNA for Use in Whole Genome Sequencing Kuiama Lewandowski,

Testing for Maternal Cell Contamination in Prenatal Samples

Application of Nuclear Magnetic Resonance to Detect Toxigenic Clostridium difficile from Stool Specimens Paul Yang, Sara Hash, Katherine Park, Charlene.

Larissa V. Furtado, Helmut C. Weigelin, Kojo S. J

Application of COLD-PCR for Improved Detection of NF2 Mosaic Mutations

Quantification of Circulating miRNAs in Plasma

Retrospective Comparison of Nucleic Acid Sequence–Based Amplification, Real-Time PCR, and Galactomannan Test for Diagnosis of Invasive Aspergillosis

Jianbo Song, Danielle Mercer, Xiaofeng Hu, Henry Liu, Marilyn M. Li

Next-Generation Sequencing for Infectious Disease Diagnosis and Management Martina I. Lefterova, Carlos J. Suarez, Niaz Banaei, Benjamin A. Pinsky The.

Rapid and Sensitive Real-Time Polymerase Chain Reaction Method for Detection and Quantification of 3243A>G Mitochondrial Point Mutation Rinki Singh,

Low Incidence of Minor BRAF V600 Mutation-Positive Subclones in Primary and Metastatic Melanoma Determined by Sensitive and Quantitative Real-Time PCR

Beatrice S. Knudsen, April N. Allen, Dale F. McLerran, Robert L

Next-Generation Genotyping by Digital PCR to Detect and Quantify the BRAF V600E Mutation in Melanoma Biopsies Pierre-Jean Lamy, Florence Castan, Nicolas.

Novel Method for PIK3CA Mutation Analysis

Detection and Quantification of BCR-ABL1 Fusion Transcripts by Droplet Digital PCR Lawrence J. Jennings, David George, Juliann Czech, Min Yu, Loren Joseph

Use of Pyrosequencing of 16S rRNA Fragments to Differentiate between Bacteria Responsible for Neonatal Sepsis Jeanne A. Jordan, Allyson R. Butchko, Mary.

Karen Snow-Bailey, Ph.D., 1961–2006

Custom Design of a GeXP Multiplexed Assay Used to Assess Expression Profiles of Inflammatory Gene Targets in Normal Colon, Polyp, and Tumor Tissue Janice.

The Effect of Nucleic Acid Extraction Platforms and Sample Storage on the Integrity of Viral RNA for Use in Whole Genome Sequencing Kuiama Lewandowski,

Nathan D. Montgomery, Sara R. Selitsky, Nirali M. Patel, D

Improved Detection of the KIT D816V Mutation in Patients with Systemic Mastocytosis Using a Quantitative and Highly Sensitive Real-Time qPCR Assay Thomas.

Clinical Experience with the Cervista HPV HR Assay

Angiotensin I-Converting Enzyme

Monique A. Johnson, Marvin J. Yoshitomi, C. Sue Richards

Optimized Allele-Specific Real-Time PCR Assays for the Detection of Common Mutations in KRAS and BRAF Alois H. Lang, Heinz Drexel, Simone Geller-Rhomberg,

The History and Impact of Molecular Coding Changes on Coverage and Reimbursement of Molecular Diagnostic Tests Susan J. Hsiao, Mahesh M. Mansukhani,

Presentation transcript:

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 120-129 (March 2012) DOI: 10.1016/j.jmoldx.2011.10.001 Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

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 2012 14, 120-129DOI: (10.1016/j.jmoldx.2011.10.001) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

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 2012 14, 120-129DOI: (10.1016/j.jmoldx.2011.10.001) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

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 2012 14, 120-129DOI: (10.1016/j.jmoldx.2011.10.001) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

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 2012 14, 120-129DOI: (10.1016/j.jmoldx.2011.10.001) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions