Rapid colorimetric quantification of PCR-amplified DNA.
A diagnostic system for rapid colorimetric quantification of the initial amount of DNA template amplified by the polymerase chain reaction is described. The method is based on co-amplification of target DNA with a cloned DNA fragment, in which a lac operator sequence has been introduced by in vitro mutagenesis. The in vitro-amplified material is immobilized on magnetic beads using the biotin-streptavidin system, and the ratio of target DNA to cloned mutated DNA can be determined using a fusion protein consisting of the Escherichia coli LacI repressor and beta-galactosidase. This method for quantitative detection of immobilized amplified nucleic acids is well adapted for rapid automated or semi-automated assays. Here, we show that it can be used to detect and quantify Plasmodium falciparum genomic DNA in clinical samples.
Lundeberg
J
J
Royal Institute of Technology, Stockholm.
Wahlberg
J
J
Uhlén
M
M
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003124:Colorimetry; D004247:DNA / Q000032:analysis*; D016054:DNA, Protozoan / Q000032:analysis*; D006801:Humans; D008288:Malaria / Q000469:parasitology*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010963:Plasmodium falciparum / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013698:Templates, Genetic
D016054:DNA, Protozoan; D004247:DNA