Detection of human immunodeficiency virus type 1 by using the polymerase chain reaction and a time-resolved fluorescence-based hybridization assay.

Abstract:

:The polymerase chain reaction (PCR) has many potential applications in the field of nucleic acid diagnostics. In particular, it has been successfully applied to the detection of pathogens present in low copy numbers such as the human immunodeficiency virus type 1. Here we describe a time-resolved fluorescence-based hybridization assay which, combined with the PCR, offers an extremely sensitive method for the detection of nucleic acids. In this assay format, the PCR is run by standard procedures and the subsequent hybridization reaction is carried out in solution by using two oligonucleotide probes, one biotinylated and one labeled with europium (Eu3+). The sandwich hybrids are then collected onto a streptavidin-coated microtitration well, and the bound Eu3+ is measured in a time-resolved fluorometer. This assay is rapid, user friendly, and quantitative and lends itself to automation. The application of this assay to the detection of human immunodeficiency virus type 1 is described.

journal_name

J Clin Microbiol

authors

Dahlén PO,Iitiä AJ,Skagius G,Frostell A,Nunn MF,Kwiatkowski M

doi

10.1128/JCM.29.4.798-804.1991

subject

Has Abstract

pub_date

1991-04-01 00:00:00

pages

798-804

issue

4

eissn

0095-1137

issn

1098-660X

journal_volume

29

pub_type

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