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Evaluation of a digital microfluidic real-time PCR platform to detect DNA of Candida albicans in blood.

Publication ,  Journal Article
Schell, WA; Benton, JL; Smith, PB; Poore, M; Rouse, JL; Boles, DJ; Johnson, MD; Alexander, BD; Pamula, VK; Eckhardt, AE; Pollack, MG ...
Published in: Eur J Clin Microbiol Infect Dis
September 2012

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.

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Published In

Eur J Clin Microbiol Infect Dis

DOI

EISSN

1435-4373

Publication Date

September 2012

Volume

31

Issue

9

Start / End Page

2237 / 2245

Location

Germany

Related Subject Headings

  • Sensitivity and Specificity
  • Real-Time Polymerase Chain Reaction
  • Microfluidics
  • Microbiology
  • Mice
  • Humans
  • Disease Models, Animal
  • Clinical Laboratory Techniques
  • Candidemia
  • Candida albicans
 

Citation

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Schell, W. A., Benton, J. L., Smith, P. B., Poore, M., Rouse, J. L., Boles, D. J., … Mitchell, T. G. (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), 2237–2245. https://doi.org/10.1007/s10096-012-1561-6
Schell, W. A., J. L. Benton, P. B. Smith, M. Poore, J. L. Rouse, D. J. Boles, M. D. Johnson, et al. “Evaluation of a digital microfluidic real-time PCR platform to detect DNA of Candida albicans in blood.Eur J Clin Microbiol Infect Dis 31, no. 9 (September 2012): 2237–45. https://doi.org/10.1007/s10096-012-1561-6.
Schell WA, Benton JL, Smith PB, Poore M, Rouse JL, Boles DJ, et al. Evaluation of a digital microfluidic real-time PCR platform to detect DNA of Candida albicans in blood. Eur J Clin Microbiol Infect Dis. 2012 Sep;31(9):2237–45.
Schell, W. A., et al. “Evaluation of a digital microfluidic real-time PCR platform to detect DNA of Candida albicans in blood.Eur J Clin Microbiol Infect Dis, vol. 31, no. 9, Sept. 2012, pp. 2237–45. Pubmed, doi:10.1007/s10096-012-1561-6.
Schell WA, Benton JL, Smith PB, Poore M, Rouse JL, Boles DJ, Johnson MD, Alexander BD, Pamula VK, Eckhardt AE, Pollack MG, Benjamin DK, Perfect JR, Mitchell TG. Evaluation of a digital microfluidic real-time PCR platform to detect DNA of Candida albicans in blood. Eur J Clin Microbiol Infect Dis. 2012 Sep;31(9):2237–2245.
Journal cover image

Published In

Eur J Clin Microbiol Infect Dis

DOI

EISSN

1435-4373

Publication Date

September 2012

Volume

31

Issue

9

Start / End Page

2237 / 2245

Location

Germany

Related Subject Headings

  • Sensitivity and Specificity
  • Real-Time Polymerase Chain Reaction
  • Microfluidics
  • Microbiology
  • Mice
  • Humans
  • Disease Models, Animal
  • Clinical Laboratory Techniques
  • Candidemia
  • Candida albicans