Evaluation of a digital microfluidic real-time PCR platform to detect DNA of Candida albicans in blood.
Abstract
Species of Candida frequently cause life-threatening infections in neonates, transplant
and intensive care unit (ICU) patients, and others with compromised host defenses.
The successful management of systemic candidiasis depends upon early, rapid diagnosis.
Blood cultures are the standard diagnostic method, but identification requires days
and less than half of the patients are positive. These limitations may be eliminated
by using real-time polymerase chain reaction (PCR) to detect Candida DNA in the blood
specimens of patients at risk. Here, we optimized a PCR protocol to detect 5-10 yeasts
in low volumes of simulated and clinical specimens. We also used a mouse model of
systemic candidiasis and determined that candidemia is optimally detectable during
the first few days after infection. However, PCR tests are often costly, labor-intensive,
and inconvenient for routine use. To address these obstacles, we evaluated the innovative
microfluidic real-time PCR platform (Advanced Liquid Logic, Inc.), which has the potential
for full automation and rapid turnaround. Eleven and nine of 16 specimens from individual
patients with culture-proven candidemia tested positive for C. albicans DNA by conventional
and microfluidic real-time PCR, respectively, for a combined sensitivity of 94%. The
microfluidic platform offers a significant technical advance in the detection of microbial
DNA in clinical specimens.
Type
Journal articleSubject
AnimalsCandida albicans
Candidemia
Clinical Laboratory Techniques
Disease Models, Animal
Humans
Mice
Microfluidics
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
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https://hdl.handle.net/10161/11061Published Version (Please cite this version)
10.1007/s10096-012-1561-6Publication Info
Schell, WA; Benton, JL; Smith, PB; Poore, M; Rouse, JL; Boles, DJ; ... Mitchell, TG (2012). Evaluation of a digital microfluidic real-time PCR platform to detect DNA of Candida
albicans in blood. Eur J Clin Microbiol Infect Dis, 31(9). pp. 2237-2245. 10.1007/s10096-012-1561-6. Retrieved from https://hdl.handle.net/10161/11061.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.
Daniel Kelly Benjamin Jr.
Kiser-Arena Distinguished Professor
Dr. Danny Benjamin is the Principal Investigator and Chair of the National Institute
of Child Health and Human Development’s Pediatric Trials Network. The Network is responsible
for designing and leading clinical trials of off-patent medicines in children of all
ages across all therapeutic areas. The team has established, or is actively studying,
the correct dosing and safety of more than 70 of the most commonly used medicines
in children. E
Melissa DePaoli Johnson
Professor in Medicine
Prognostic indicators for patients with Candida spp. bloodstream infections Antifungal
pharmacokinetics/pharmacodynamics Immunogenetics among patients with candidiasis Management
of the HIV infected patient and antiretroviral pharmacotherapy Antibacterial drug
utilization, resistance, and appropriate prescribingAntimicrobial Stewardship
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
Phillip Brian Smith
Samuel L. Katz Distinguished Professor of Pediatrics
Dr. Smith completed his residency in pediatrics and a fellowship in neonatal medicine
at Duke University Medical Center in 2004 and 2007, respectively. He completed an
MHS in clinical research from Duke University in 2006 and an MPH in biostatistics
from the University of North Carolina at Chapel Hill in 2009. His research is focused
on pediatric drug safety, neonatal pharmacology, and the epidemiology of neonatal
infections. Dr. Smith is or has been the protocol chair for more than 14 studie
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