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PCR-based diagnosis of Naegleria sp. infection in formalin-fixed and paraffin-embedded brain sections
Marc Schild1, Christian Gianinazzi, Bruno Gottstein
1Institute of Parasitology, University of Berne, Länggass-Strasse 122, CH-3012 Bern, Switzerland.
Abstract:
We developed a real-time PCR which allowed the highly sensitive detection of Naegleria fowleri in histological brain tissue sections from experimentally infected mice. This genus-specific small-subunit (18S) rRNA gene-based PCR can complement conventional (immuno-) histology for the diagnosis of primary amoebic meningoencephalitis in paraffin-embedded brain necropsy specimens that had been fixed in formalin buffered with phosphate-buffered saline.
Insights
We created a sensitive real-time PCR test for detecting Naegleria fowleri in mouse brain tissue. This molecular method aids in diagnosing primary amoebic meningoencephalitis from preserved samples.
Area of Science:
- Molecular Biology
- Pathology
- Parasitology
Background:
- Primary amoebic meningoencephalitis (PAM) is a rare but severe infection.
- Diagnosis often relies on traditional histological methods.
- Naegleria fowleri is the causative agent of PAM.
Purpose of the Study:
- To develop a highly sensitive molecular diagnostic tool for Naegleria fowleri.
- To evaluate the utility of real-time PCR in detecting the pathogen in brain tissue.
Main Methods:
- Development of a genus-specific real-time PCR assay targeting the 18S rRNA gene.
- Application of the PCR assay to histological brain tissue sections from experimentally infected mice.
- Comparison with conventional histological techniques.
Main Results:
- The developed real-time PCR demonstrated high sensitivity for Naegleria fowleri detection.
- The assay successfully identified the pathogen in paraffin-embedded, formalin-fixed brain tissue.
- The PCR method complements existing diagnostic approaches.
Conclusions:
- Real-time PCR offers a sensitive and specific method for Naegleria fowleri detection.
- This molecular technique is valuable for diagnosing PAM in post-mortem specimens.
- The assay can aid in confirming infections when conventional methods are limited.
