Microfluidic platform versus conventional real-time polymerase chain reaction for the detection of Mycoplasma pneumoniae in respiratory specimens.
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.
School of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
Elizabeth
E
Wulff-Burchfield
Wiley A
WA
Schell
Allen E
AE
Eckhardt
Michael G
MG
Pollack
Zhishan
Z
Hua
Jeremy L
JL
Rouse
Vamsee K
VK
Pamula
Vijay
V
Srinivasan
Jonathan L
JL
Benton
Barbara D
BD
Alexander
David A
DA
Wilfret
Monica
M
Kraft
Charles B
CB
Cairns
John R
JR
Perfect
Thomas G
TG
Mitchell
D001331:Automation; D001431:Bacteriological Techniques / Q000379:methods*; D017714:Community-Acquired Infections / Q000382:microbiology; D006801:Humans; D044085:Microfluidics / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009177:Mycoplasma pneumoniae / Q000235:genetics / Q000302:isolation & purification*; D009305:Nasopharynx / Q000382:microbiology; D011019:Pneumonia, Mycoplasma / Q000175:diagnosis* / Q000382:microbiology; D019095:Point-of-Care Systems; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors