Colorimetric detection for PCR amplified HIV-1 DNA using magnetic beads.
A rapid and nonradioactive detection method for polymerase chain reaction (PCR) amplified HIV-1 DNA was developed using a colorimetric detection system. Hybridization between biotin-labeled amplified targets and digoxigenin-capture probes occurs in solution followed by efficient and rapid capture onto streptavidin-magnetic beads. The presence of the digoxigenin-capture probe hybridized with biotin-labeled targets is then detected by antidigoxigenin-alkaline phosphatase conjugates using a colorimetric substrate. This approach is highly sensitive and can detect less than 10 HIV targets prior to PCR in approximately 50 min.
Suzuki
K
K
Biomedical Research Center, Olympus Corporation, East Setauket, NY 11733.
Okamoto
N
N
Kano
T
T
D001426:Bacterial Proteins; D001483:Base Sequence; D001710:Biotin; D003124:Colorimetry / Q000379:methods*; D015342:DNA Probes; D004277:DNA, Single-Stranded / Q000737:chemistry; D004279:DNA, Viral / Q000302:isolation & purification*; D004076:Digoxigenin; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008280:Magnetics; D008863:Microspheres; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019809:Streptavidin; D013997:Time Factors
D001426:Bacterial Proteins; D015342:DNA Probes; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D001710:Biotin; D019809:Streptavidin; D004076:Digoxigenin