Nucleic acids, detection with

Devries, E.F.A., Schasfoort, R.B.M., Vanderplas,)., and Greve,). (1994) Nucleic acid detection with surface plasmon resonance using cationic latex. Biosensors i Biodectronics, 9 (7),... 

Cunningham, M. W. (1991) Nucleic acid detection with light. Life Sci. 6,2-5. 

Ogul chansky TYu, Yashchuk VM, Losytskyy MYu, Kocheshev IO, Yarmoluk SM (2000) Interaction of cyanine dyes with nucleic acids. XVII. Towards an aggregation of cyanine dyes in solutions as a factor facilitating nucleic acid detection. Spectrochim Acta A 56 805-814... 

Dioxetanes, labeled with triggers sensitive to the alkaline-phosphatase enzyme, serve as highly sensitive chemiluminescent probes in numerous bioassays. Current applications include immunoassays, membrane-based detection of proteins and nucleic acids, and microplate-based and array-based nucleic-acid detection. ... 

Multiple in situ nucleic acid detection can be achieved in two ways- by sequential hybridization with probes labeled with the same reporter and by simultaneous hybridization using probes labeled with different reporters. Any of the reporters listed above can be combined to allow dual nucleic acid detection, but we have found biotm and digoxigenin to be the most generally useful, since they are safe and sensitive Probes labeled with these reporters can be combined m NISH and detected differentially according to the scheme presented m Fig. 1 (6) 2. Pretreatment of cells/tissues Having chosen the reporter and incorporated it into a probe, the cell/tissue to be analyzed is prepared for hybridization The following requirements must be met to allow a successful hybridization reaction to take place. 

Fig. 1. Dual nucleic acid detection. The target DNAs (A and B) are denatured and hybridized with complementary DNA probes labeled with biotin (Bio ), or digoxigenm (Dig ). Biotin and digoxigenm residues are detected, respectively, with avidin peroxidase (red) and antibody to digoxigenin labeled with alkaline phosphatase (blue).

Detection of the sites of hybridization-dependent binding of biotinylated probe to the filters is most readily conducted with commercially available kits. Favorable results have been obtained with the BluGene Nonradioactive Nucleic Acid Detection System from BRL. Follow the manufacturer s instructions when carrying out the following steps. After washing, sequentially expose the filters to streptavidin and biotinylated alkaline phosphatase (or to a conjugate of these two proteins) This causes the immobilization of alkaline phosphatase at sites of positive hybridization... 

P.M. Armistead and H.H. Thorp, Modification of indium tin oxide electrodes with nucleic acids detection of attomole quantities of immobilized DNA by electrocatalysis, Anal. Chem., 72 (2000) 3764-3770. 

This method provided a simple and fast procedure to identify the presence of a nucleic acid sequence with good sensitivity. On average, the liposomes entrapped several thousand dye molecules, which provided intrinsic amplification of individual binding events. A qualitative measure of nucleic acid hybridization by visualization of the colour on the dip stick was intrinsically provided and quantitative measurements were also possible by use of a reflectometer. This approach resulted in a limit of detection of 1 fmol and a dynamic range of over two orders of magnitude. Maximum signal-to-noise was obtained with 0.2 mol% of probe sequence immobilized on the liposomes. 

It is also possible to incubate radioactive or fluorescent nucleic acid probes with sections of tissues or even chromosomes, wash away excess probe and then detect where the probe has hybridized. This technique (in situ hybridization) has proved to be very powerful in determining which cells in a complex tissue such as the mammalian brain express a particular gene and for locating specific genes on individual chromosomes. 

The greatest advances in the detection of chlamydial infection involved the development of various nucleic acid detection methods. Similar to EIA, the DNA hybridization probe test is easy to perform, and a large number of samples can be processed at the same time. Overall, the sensitivity and specificity of the DNA probe tests are greater than with EIA. " " ... 

---