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Rapid detection of toxoplasmic nucleic acid by enzyme‐linked immunofiltration‐assay after membrane transfer
Author(s) -
Aubert Dominique,
Lepan Herve,
Lemaire Pascale,
Foudrinier Frederique,
MarxChemla Cathy,
Bonhomme Annie,
Pi JeanMichel
Publication year - 1995
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150160160
Subject(s) - nitrocellulose , chromatography , reagent , membrane , chemistry , reproducibility , repeatability , nucleic acid , biochemistry
We report the use of enzyme‐linked immunofiltration‐assay (ELIFA) for rapid detection of Toxoplasma gondii DNA after transfer to nitrocellulose or nylon membranes. Saturation, specific labeling with a nonradioactive oligonucleotide probe and washing steps are included in an active filtration process, which is controled by microcomputer regulating reagent flow. This method provides excellent specificity, reproducibility and repeatability. Recycling in a closed circuit or by reversing the direction of flow provides the advantage of reducing the required volume of costly reagents ( e.g. probes). ELIFA does not require lengthy membrane manipulation, and hybridization as well as stringency and revelation steps can be automated. The procedure can be carried out in less than 2 h, which is a major time saving compared to conventional solid‐phase molecular hybridization techniques.