Accelerated detection of DNA on membranes by automated enzyme-linked immunofiltration assay

被引：16

作者：

Aubert, D ^{[1
]}

Toubas, D ^{[1
]}

Foudrinier, F ^{[1
]}

Villena, I ^{[1
]}

Gomez, JE ^{[1
]}

Marx, C ^{[1
]}

Lepan, H ^{[1
]}

Lemaire, P ^{[1
]}

Jacquier, P ^{[1
]}

Bonhomme, A ^{[1
]}

Pinon, JM ^{[1
]}

机构：

[1] UNIV BERN, INST PARASITOL, CH-3001 BERN, SWITZERLAND

来源：

ANALYTICAL BIOCHEMISTRY | 1997年 / 247卷 / 01期

关键词：

D O I：

10.1006/abio.1997.2030

中图分类号：

Q5 [生物化学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

Methods used to detect DNA after transfer to nitrocellulose or nylon membranes are all based on slow incubation with agitation. We describe an application of the ELIFA technique (enzyme-linked immunofiltration assay) for rapid detection of DNA immobilized on a membrane by active filtration of the reagents across the membrane. The different steps (saturation, hybridization to a nonisotopically labeled probe, washing, and immunoenzymatic revelation) are automated and controlled by a microcomputer that determines the direction of how and how rates of the solutions through the membrane. We applied this method to the detection of Toxoplasma gondii DNA in 108 samples of amniotic fluid during antenatal tests for toxoplasmosis and compared the results with those obtained by the conventional method. In addition to a major time saving (2 h against almost 15 h), automation improves reproducibility and avoids manipulation of the membranes between the different steps, while keeping the same sensitivity and specificity as the standard method. (C) 1997 Academic Press.

引用

页码：25 / 29

页数：5

共 23 条

[1]

AMSELEM S, 1988, AM J HUM GENET, V43, P95

[2]

RAPID DETECTION OF TOXOPLASMIC NUCLEIC-ACID BY ENZYME-LINKED IMMUNOFILTRATION-ASSAY AFTER MEMBRANE TRANSFER [J].