Fluorometric broad-range screening of compounds with affinity for nucleic acids.
The potential of a nucleic acid-based optical bioprobe for environmental measurements and drug monitoring is described. The sensor employs the long-wavelength intercalating fluorophore TO-PRO-3 (TP3). Compounds that interact with the TP3-DNA complex are indirectly detected by a decrease in the fluorescence intensity. We found that the configuration and length of the DNA dramatically affected the intensity of the fluorescence emitted from the TP3-DNA complex. We compared nucleic acids from different sources and optimized the system for pBR322 plasmid DNA (4363 bp) digested by HindIII restriction endonuclease. This endonuclease has a single recognition site in plasmid pBR322. In the proposed method, we attempt to combine broad-range detection with rapid and simple operation. A fiber-optic capillary fluorescence system was used to analyze toxic aromatic amines, antibiotics, and several kinds of antitumor drugs, using small amounts of sample, down to 10 muL, with a sensitivity comparable to that of current electrochemical methods. The detection limit can be as low as a few ppb or submicromolar. This approach is useful for routine screening in environmental monitoring or for controlling cytotoxic drug administration. The ease of operation and the rapid response allow high-throughput screening.
Liu
Yang
Y
Pure and Applied Biochemistry, Lund University, P.O. Box 124, SE-22100 Lund, Sweden.
Danielsson
Bengt
B
D000588:Amines / Q000032:analysis; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002232:Carbocyanines / Q000737:chemistry / Q000378:metabolism; D002701:Chloramphenicol / Q000032:analysis; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D004784:Environmental Monitoring / Q000379:methods; D005336:Fiber Optic Technology / Q000295:instrumentation / Q000379:methods; D005453:Fluorescence; D005470:Fluorometry / Q000295:instrumentation / Q000379:methods*; D008970:Molecular Weight; D009696:Nucleic Acids / Q000378:metabolism*; D055100:Optical Fibers; D010957:Plasmids / Q000235:genetics; D012680:Sensitivity and Specificity; D013997:Time Factors
D000588:Amines; D002232:Carbocyanines; D009696:Nucleic Acids; C098830:TO-PRO-3; D002701:Chloramphenicol; D004247:DNA; D015247:Deoxyribonuclease HindIII