Microwave irradiation-accelerated in situ hybridization technique for HIV detection.
High frequency irradiation generated in a common household microwave oven was used to establish an in situ hybridization technique for rapid detection of HIV sequences in infected cells. A biotin-labeled DNA probe was subsequently detected either by an alkaline phosphatase-based colorimetric reaction or by fluorescence. When compared to standard hybridization procedures with radioactive or nonradioactive probes, microwave energy-mediated hybridization results in equal sensitivity and diminished background. The main advantage of this method, however, is the drastic reduction in time, allowing completion of the whole procedure, from sample preparation to hybrid signal visualization, within one hour. In addition to HIV detection, the approach described can be applied for the diagnosis of other viral infections and may stimulate the development of nucleic acid hybridization techniques based on microwave irradiation.
Bourinbaiar
A S
AS
Population Council, Center for Biomedical Research, New York, NY.
Zacharopoulos
V R
VR
Phillips
D M
DM
D001710:Biotin; D002478:Cells, Cultured; D015342:DNA Probes; D004279:DNA, Viral / Q000302:isolation & purification; D006678:HIV / Q000737:chemistry*; D015658:HIV Infections / Q000175:diagnosis*; D006651:Histocytochemistry / Q000379:methods*; D006801:Humans; D008872:Microwaves; D009693:Nucleic Acid Hybridization
D015342:DNA Probes; D004279:DNA, Viral; D001710:Biotin