Microfluidic platform versus conventional real-time polymerase chain reaction for the detection of Mycoplasma pneumoniae in respiratory specimens.
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 articleSubject
AutomationBacteriological 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
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https://hdl.handle.net/10161/11070Published Version (Please cite this version)
10.1016/j.diagmicrobio.2009.12.020Publication 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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Show full item recordScholars@Duke
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.
Monica Kraft
Adjunct Professor in the Department of Medicine
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
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
Wiley Alexander Schell
Associate Professor Emeritus in Medicine
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