Polymerase chain reaction to detect the pathogenic Naegleria fowleri: application to water samples

O. Sparagano, E. Drouet, G. Denoyel, P. Pernin

Research output: Contribution to journalArticle

Abstract

Search of Naegleria fowleri in water samples was performed by using polymerase chain reaction (PCR) and hybridization with an internal digoxigenin labeled probe. First, the specificity of our technique was tested on axenic or monoxenic cultures. Each of the 13 strains originated from various organisms were not. The sensitivity with 40 amplification cycles was a single cell (cyst or trophozoite). Secondly, we have several protocols of cell recovery (i.e. sonication, freeze-thaw cycles, formamide, glass beads and chemical lysis) on artificially contaminated tap water samples. We showed that sonication, freeze-thaw cycles and formamide are better than chemical lysis and glass beads with proteinase K. Work is in progress to use this technique on environmental samples.

Original languageEnglish
Pages (from-to)81-88
Number of pages8
JournalJournal of Microbiological Methods
Volume19
Issue number2
DOIs
Publication statusPublished - 1 Jan 1994
Externally publishedYes

Fingerprint

Naegleria fowleri
Sonication
Glass
Digoxigenin
Endopeptidase K
Polymerase Chain Reaction
Trophozoites
Water
Cysts
formamide

Keywords

  • Amoeba
  • Cellular lysis
  • Polymerase chain reaction
  • Water sample

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Microbiology (medical)

Cite this

Polymerase chain reaction to detect the pathogenic Naegleria fowleri : application to water samples. / Sparagano, O.; Drouet, E.; Denoyel, G.; Pernin, P.

In: Journal of Microbiological Methods, Vol. 19, No. 2, 01.01.1994, p. 81-88.

Research output: Contribution to journalArticle

Sparagano, O. ; Drouet, E. ; Denoyel, G. ; Pernin, P. / Polymerase chain reaction to detect the pathogenic Naegleria fowleri : application to water samples. In: Journal of Microbiological Methods. 1994 ; Vol. 19, No. 2. pp. 81-88.
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