Current Microbiology

, Volume 53, Issue 6, pp 506–509

Quantitative Detection and Differentiation of Free-Living Amoeba Species Using SYBR Green–Based Real-Time PCR Melting Curve Analysis

  • Jonas Behets
  • Priscilla Declerck
  • Yasmine Delaedt
  • Lieve Verelst
  • Frans Ollevier
Article

DOI: 10.1007/s00284-006-0241-0

Cite this article as:
Behets, J., Declerck, P., Delaedt, Y. et al. Curr Microbiol (2006) 53: 506. doi:10.1007/s00284-006-0241-0

Abstract

Real-time polymerase chain reaction melting curve analysis (MCA) allows differentiation of several free-living amoebae species. Distinctive characteristics were found for Naegleria fowleri, N. lovaniensis, N. australiensis, N. gruberi, Hartmanella vermiformis, and Willaertia magna. Species specificity of the amplicons was confirmed using agarose gel electrophoresis and sequence-based approaches. Amplification efficiency ranged from 91% to 98%, indicating the quantitative potential of the assay. This MCA approach can be used for quantitative detection of free-living amoebae after cultivation but also as a culture-independent detection method.

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Jonas Behets
    • 1
  • Priscilla Declerck
    • 1
  • Yasmine Delaedt
    • 1
  • Lieve Verelst
    • 2
  • Frans Ollevier
    • 1
  1. 1.Laboratory of Aquatic EcologyKatholieke Universiteit Leuven (KUL)LeuvenBelgium
  2. 2.LABORELECLinkebeekBelgium

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