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

Elizabeth Wulff-Burchfield, Wiley A. Schell, Allen E. Eckhardt, Michael G. Pollack, Zhishan Hua, Jeremy L. Rouse, Vamsee K. Pamula, Vijay Srinivasan, Jonathan L. Benton, Barbara D. Alexander, David A. Wilfret, Monica Kraft, Charles B. Cairns, John R. Perfect, Thomas G. Mitchell

Research output: Contribution to journalArticle

30 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)22-29
Number of pages8
JournalDiagnostic Microbiology and Infectious Disease
Volume67
Issue number1
DOIs
StatePublished - May 1 2010
Externally publishedYes

Keywords

  • Community-acquired pneumonia
  • DNA-based diagnostics
  • Diagnostic microbiology
  • Mycoplasma pneumoniae
  • Real-time PCR

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases

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  • Cite this

    Wulff-Burchfield, E., Schell, W. A., Eckhardt, A. E., Pollack, M. G., Hua, Z., Rouse, J. L., Pamula, V. K., Srinivasan, V., Benton, J. L., Alexander, B. D., Wilfret, D. A., Kraft, M., Cairns, C. B., Perfect, J. R., & Mitchell, T. G. (2010). Microfluidic platform versus conventional real-time polymerase chain reaction for the detection of Mycoplasma pneumoniae in respiratory specimens. Diagnostic Microbiology and Infectious Disease, 67(1), 22-29. https://doi.org/10.1016/j.diagmicrobio.2009.12.020