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Exp Parasitol. 2010 Sep;126(1):37-41. doi: 10.1016/j.exppara.2009.11.001. Epub 2009 Nov 15.

A real-time PCR diagnostic method for detection of Naegleria fowleri.

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  • 1Regional Authority of Public Health Banská Bystrica, Department of Molecular Biology, Cesta k nemocnici 1, Banská Bystrica 975 56, Slovakia. lucia.madarova@vzbb.sk

Abstract

Naegleria fowleri is a free-living amoeba that can cause primary amoebic meningoencephalitis (PAM). While, traditional methods for diagnosing PAM still rely on culture, more current laboratory diagnoses exist based on conventional PCR methods; however, only a few real-time PCR processes have been described as yet. Here, we describe a real-time PCR-based diagnostic method using hybridization fluorescent labelled probes, with a LightCycler instrument and accompanying software (Roche), targeting the Naegleria fowleriMp2Cl5 gene sequence. Using this method, no cross reactivity with other tested epidemiologically relevant prokaryotic and eukaryotic organisms was found. The reaction detection limit was 1 copy of the Mp2Cl5 DNA sequence. This assay could become useful in the rapid laboratory diagnostic assessment of the presence or absence of Naegleria fowleri.

Copyright 2009 Elsevier Inc. All rights reserved.

PMID:
19919836
[PubMed - indexed for MEDLINE]
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