Comparison of real-time PCR methods for the detection of Naegleria fowleri in surface water and sediment
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Naegleria fowleri is a thermophilic free-living ameba found in freshwater environments worldwide. It is the cause of a rare but potentially fatal disease in humans known as primary amebic meningoencephalitis. Established N. fowleri detection methods rely on conventional culture techniques and morphological examination followed by molecular testing. Multiple alternative real-time PCR assays have been published for rapid detection of Naegleria spp. and N. fowleri. Foursuch assays were evaluated for the detection of N. fowleri from surface water and sediment. The assays were compared for thermodynamic stability, analytical sensitivity and specificity, detection limits, humic acid inhibition effects, and performance with seeded environmental matrices. Twenty-one ameba isolates were included in the DNA panel used for analytical sensitivity and specificity analyses. N. fowleri genotypes I and III were used for method performance testing. Two of the real-time PCR assays were determined to yield similar performance data for specificity and sensitivity for detecting N. fowleri in environmental matrices.
KeywordsNaegleria fowleri Real-time PCR Environmental detection Method comparison
We would like to thank Dr. Jothikumar Narayanan (CDC) for his guidance on implementing the real-time PCR assays for this study and evaluation of study data. The findings and conclusions in this presentation are those of the authors and do not necessarily represent those of the Centers for Disease Control and Prevention and Emory University. The use of trade names and names of commercial sources is for identification only and does not imply endorsement by the Centers for Disease Control and Prevention or the US Department of Health and Human Services.
- De Jonckheere JF (2002) A century of research on the amoeboflagellate genus Naegleria. Acta Protozool 41(4):309–342Google Scholar
- HillVR, Jothikumar N, Vinje J, Cromeans TL (2010) Sample Preparation Methods for Molecular Techniques for Drinking Water. Project Report #3108, Water Research FoundationGoogle Scholar
- Mull B, Narayanan J, Hill VR (2013) Improved method for the detection and quantification of Naegleria fowleri in water and sediment using immunomagnetic separation and real-time PCR. J Parasitol Res 2013Google Scholar
- Natural Resources Conservation Service (2013) Distribution Maps of Dominant Soil Orders. Accessed 11/06/2013, from http://soils.usda.gov/technical/classification/orders/
- Yoder JS, Straif-Bourgeois S, Roy SL, Moore TA, Visvesvara GS, Ratard RC, Hill VR, Wilson JD, Linscott AJ, Crager R, Kozak NA, Sriram R, Narayanan J, Mull B, Kahler AM, Schneeberger C, da Silva AJ, Poudel M, Baumgarten KL, Xiao L, Beach MJ (2012) Primary amebic meningoencephalitis deaths associated with sinus irrigation using contaminated tap water. Clin Infect Dis 55(9):e79–e85CrossRefPubMedGoogle Scholar