Rapid colorimetric detection of in vitro amplified DNA sequences.
A colorimetric assay to detect immobilized amplified nucleic acids has been designed. This approach provides a rapid assay, suitable for clinical diagnosis, to analyze DNA sequences amplified by the polymerase chain reaction. The specific DNA sequences are captured on a solid support by the use of a recombinant fusion protein consisting of the Escherichia coli lac repressor and staphylococcal protein A. The biotin streptavidin system is used to detect the immobilized material. Positive samples can be analyzed by direct solid-phase sequencing. Here, we show that this nonradioactive concept can be used for analysis of Staphylococci and Streptococci and for specific detection of the protozoa Plasmodium falciparum in clinical samples.
Royal Institute of Technology, Department of Biochemistry and Biotechnology, Stockholm, Sweden.
J
J
Lundeberg
J
J
Wahlberg
M
M
Holmberg
U
U
Pettersson
M
M
Uhlén
D000818:Animals; D001483:Base Sequence; D003124:Colorimetry / Q000379:methods; D004247:DNA / Q000032:analysis* / Q000097:blood; D004269:DNA, Bacterial / Q000032:analysis; D005821:Genetic Techniques; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D010963:Plasmodium falciparum / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D013210:Staphylococcus / Q000235:genetics; D013291:Streptococcus / Q000235:genetics
D004269:DNA, Bacterial; D004247:DNA