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.
Wulff-Burchfield
Elizabeth
E
School of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
Schell
Wiley A
WA
Eckhardt
Allen E
AE
Pollack
Michael G
MG
Hua
Zhishan
Z
Rouse
Jeremy L
JL
Pamula
Vamsee K
VK
Srinivasan
Vijay
V
Benton
Jonathan L
JL
Alexander
Barbara D
BD
Wilfret
David A
DA
Kraft
Monica
M
Cairns
Charles B
CB
Perfect
John R
JR
Mitchell
Thomas G
TG
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