Differentiation of Naegleria fowleri and other naegleriae by polymerase chain reaction and hybridization methods

Olivier Sparagano

Research output: Contribution to journalArticle

Abstract

In order to detect and identify Naegleria fowleri strains an assay based on the Polymerase Chain Reaction (PCR) was evaluated. The amplified DNA fragments were detected by gel electrophoresis and ethidium bromide staining, followed by Southern blot hybridization with an internal digoxigenin-labeled probe. A set of primers (B1B2) which flank a 678-bp region within a virulence-associated gene, allowed for the highly specific identification of N. fowleri, since Naegleriae (N. lovaniensis, N. australiensis, N. gruberi, N. andersoni and N. jadini) and other Protozoa did not react. These primers did not detect amplification products from various organisms: Gram-positive bacteria, algae, y, yeasts and human DNA. Whereas a second set of primers (A1A2), which flank a different sequence, detected various Naegleriae and Acanthamoebae strains. After 40 amplification cycles, the limit of detection was a single cell (cyst or trophozoite). Thus, the PCR appears to be a rapid and powerful tool for identification and detection of N. fowleri.

Original languageEnglish
Pages (from-to)325-330
Number of pages6
JournalFEMS Microbiology Letters
Volume110
Issue number3
DOIs
Publication statusPublished - 1 Jul 1993
Externally publishedYes

Fingerprint

Naegleria fowleri
Polymerase Chain Reaction
Acanthamoeba
Digoxigenin
Trophozoites
Ethidium
DNA
Gram-Positive Bacteria
Southern Blotting
Virulence
Electrophoresis
Limit of Detection
Cysts
Yeasts
Gels
Staining and Labeling
Genes

Keywords

  • DNA probe
  • Naegleria fowleri
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Genetics

Cite this

Differentiation of Naegleria fowleri and other naegleriae by polymerase chain reaction and hybridization methods. / Sparagano, Olivier.

In: FEMS Microbiology Letters, Vol. 110, No. 3, 01.07.1993, p. 325-330.

Research output: Contribution to journalArticle

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