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Microfluidic platform versus conventional real-time polymerase chain reaction for the detection of Mycoplasma pneumoniae in respiratory specimens.

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Date
2010-05
Authors
Wulff-Burchfield, Elizabeth
Schell, Wiley A
Eckhardt, Allen E
Pollack, Michael G
Hua, Zhishan
Rouse, Jeremy L
Pamula, Vamsee K
Srinivasan, Vijay
Benton, Jonathan L
Alexander, Barbara D
Wilfret, David A
Kraft, Monica
Cairns, Charles B
Perfect, John R
Mitchell, Thomas G
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(15 total)
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Abstract
Rapid, accurate diagnosis of community-acquired pneumonia (CAP) due to Mycoplasma pneumoniae is compromised by low sensitivity of culture and serology. Polymerase chain reaction (PCR) has emerged as a sensitive method to detect M. pneumoniae DNA in clinical specimens. However, conventional real-time PCR is not cost-effective for routine or outpatient implementation. Here, we evaluate a novel microfluidic real-time PCR platform (Advanced Liquid Logic, Research Triangle Park, NC) that is rapid, portable, and fully automated. We enrolled patients with CAP and extracted DNA from nasopharyngeal wash (NPW) specimens using a biotinylated capture probe and streptavidin-coupled magnetic beads. Each extract was tested for M. pneumoniae-specific DNA by real-time PCR on both conventional and microfluidic platforms using Taqman probe and primers. Three of 59 (5.0%) NPWs were positive, and agreement between the methods was 98%. The microfluidic platform was equally sensitive but 3 times faster and offers an inexpensive and convenient diagnostic test for microbial DNA.
Type
Journal article
Subject
Automation
Bacteriological Techniques
Community-Acquired Infections
Humans
Microfluidics
Molecular Diagnostic Techniques
Mycoplasma pneumoniae
Nasopharynx
Pneumonia, Mycoplasma
Point-of-Care Systems
Polymerase Chain Reaction
Sensitivity and Specificity
Time Factors
Permalink
https://hdl.handle.net/10161/11070
Published Version (Please cite this version)
10.1016/j.diagmicrobio.2009.12.020
Publication Info
Wulff-Burchfield, Elizabeth; Schell, Wiley A; Eckhardt, Allen E; Pollack, Michael G; Hua, Zhishan; Rouse, Jeremy L; ... Mitchell, Thomas G (2010). Microfluidic platform versus conventional real-time polymerase chain reaction for the detection of Mycoplasma pneumoniae in respiratory specimens. Diagn Microbiol Infect Dis, 67(1). pp. 22-29. 10.1016/j.diagmicrobio.2009.12.020. Retrieved from https://hdl.handle.net/10161/11070.
This is constructed from limited available data and may be imprecise. To cite this article, please review & use the official citation provided by the journal.
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Scholars@Duke

Alexander

Barbara Dudley Alexander

Professor of Medicine
Clinical research related to infectious complications of solid organ and bone marrow transplantation, with a particular interest in the treatment and rapid diagnosis of fungal disease. Training the next generation of Transplant Infectious Disease Physicians is a special focus of mine as the Principal Investigator of our Interdisciplinary T32 Training Program funded the NIH. 
Kraft

Monica Kraft

Adjunct Professor in the Department of Medicine
Mitchell

Thomas Greenfield Mitchell

Associate Professor Emeritus in Molecular Genetics and Microbiology
Among patients with AIDS, leukemia or other cancers, organ or bone marrow transplants, and similar immunocompromising risk factors, the incidence of opportunistic mycoses and the number of different fungal pathogens are increasing dramatically. For many of these fungi, the definition of a species and the recognition of pathogen are highly problematic. Conventional methods of identification are based on morphological and physiological characteristics and are often time-consuming, difficult to int
Perfect

John Robert Perfect

James B. Duke Distinguished Professor of Medicine
Research in my laboratory focuses around several aspects of medical mycology. We are investigating antifungal agents (new and old) in animal models of candida and cryptococcal infections. We have examined clinical correlation of in vitro antifungal susceptibility testing and with in vivo outcome. Our basic science project examines the molecular pathogenesis of cryptococcal infections. We have developed a molecular foundation for C. neoformans, including transformation systems, gene disr
Schell

Wiley Alexander Schell

Associate Professor Emeritus in Medicine
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