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. 2006 Dec;53(6):506-9.
doi: 10.1007/s00284-006-0241-0. Epub 2006 Nov 13.

Quantitative detection and differentiation of free-living amoeba species using SYBR green-based real-time PCR melting curve analysis

Jonas Behets  1 , Priscilla Declerck, Yasmine Delaedt, Lieve Verelst, Frans Ollevier
Affiliations

Quantitative detection and differentiation of free-living amoeba species using SYBR green-based real-time PCR melting curve analysis

Jonas Behets et al. Curr Microbiol. 2006 Dec.

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.

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