Rapid detection of toxoplasmic nucleic acid by enzyme‐linked immunofiltration‐assay after membrane transfer
- 1 January 1995
- journal article
- Published by Wiley in Electrophoresis
- Vol. 16 (1), 354-356
- https://doi.org/10.1002/elps.1150160160
Abstract
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.Keywords
This publication has 18 references indexed in Scilit:
- Prenatal Diagnosis of Congenital Toxoplasmosis with a Polymerase-Chain-Reaction Test on Amniotic FluidNew England Journal of Medicine, 1994
- Contribution of a New PCR assay to the prenatal diagnosis of congenital toxoplasmosisPrenatal Diagnosis, 1992
- Nonradioactive labeling of oligonucleotides in vitro with the hapten digoxigenin by tailing with terminal transferaseAnalytical Biochemistry, 1991
- Rapid detection of proteins by enzyme‐linked immunofiltration assay after transfer onto nitrocellulose membranesElectrophoresis, 1990
- The use of biotinylated DNA probes in parentage testing: Non‐isotopic labeling and non‐toxic extractionElectrophoresis, 1988
- Enzymatic Amplification of β-Globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell AnemiaScience, 1985
- An enzyme-linked immuno-filtration assay used to compare infant and maternal antibody profiles in toxoplasmosisJournal of Immunological Methods, 1985
- Detection of Enterotoxigenic Escherichia coli by DNA Colony HybridizationThe Journal of Infectious Diseases, 1980
- Detection of specific sequences among DNA fragments separated by gel electrophoresisJournal of Molecular Biology, 1975
- A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membraneJournal of Molecular Biology, 1965