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

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

United States

2015-12-03T21:07:25Z

2010-05

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.

http://www.ncbi.nlm.nih.gov/pubmed/20227222

S0732-8893(09)00511-2

1879-0070

https://hdl.handle.net/10161/11070

eng

Elsevier BV

Diagn Microbiol Infect Dis

10.1016/j.diagmicrobio.2009.12.020

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

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

Journal article

Alexander, Barbara D|0000-0001-5868-0529

Perfect, John R|0000-0002-6606-9460|0000-0003-3465-5518

http://www.ncbi.nlm.nih.gov/pubmed/20227222

22

29

1

Basic Science Departments

Clinical Science Departments

Duke

Medicine

Medicine, Infectious Diseases

Medicine, Pulmonary, Allergy, and Critical Care Medicine

Molecular Genetics and Microbiology

Pathology

School of Medicine

Published

67