Nucleic acid detection

Many nucleic acid detection strategies use target amplification, signal amplification or both. Invader, branched DNA (bDNA) and rolling circle amplification (RCA) are three approaches. 

Chemiluminescence reactions are currently exploited mainly either for analyte concentration measurements or for immunoanalysis and nucleic acid detection. In the latter case, a compound involved in the light emitting reaction is used as a label for immunoassays or for nucleic acid probes. In the former case, the analyte of interest directly participates in a chemiluminescence reaction or undergoes a chemical or an enzymatic transformation in such a way that one of the reaction products is a coreactant of a chemiluminescence reaction. In this respect, chemiluminescent systems that require H2O2 for the light emission are of particular interest in biochemical analysis. Hydrogen peroxide is in fact a product of several enzymatic reactions, which can be then coupled to a chemiluminescent detection. 

Kricka, L.J. (1999) Nucleic acid detection technologies—labels, strategies, and formats. Clinical Chemistry 45(4), 453-458. 

F. Uslu, S. Ingebrandt, D. Mayer, S. Bocker-Meffert, M. Odenthal, and A. Offenhausser, Labelfree fully electronic nucleic acid detection system based on a field-effect transistor device. Biosens. Bioelectron. 19, 1723-1731 (2004). 

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... 

Yang X, Zhao X, Zuo X, Wang K, Wen J, Zhang H (2009) Nucleic acids detection using cationic fluorescent polymer based on one-dimensional microfluidic beads array. Talanta 77 1027-1031... 

Examples of its use include protein and nucleic acid detection, enzyme-labelled fluorescence, in the intrinsic fluorescence of normal and cancer cells, as external... 

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. ... 

Mansfield, E. S. Worley, J. M. McKenzie, S. E. Surrey, S. Rappaport, E. Fortina, P. Nucleic acid detection using non-radioactive labelling methods. Mol. Cell. Probes 1995, 5(3), 145-156. 

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).

Reagents for the detection of filter-bound biotin are obtained from BRL (BlueGene Nonradioactive Nucleic Acid Detection System) Comparable materials are available from Bio-Rad Laboratories... 

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. 

M. Dequaire and A. Heller, Screen-printing of nucleic acid detecting carbon electrodes, Anal. Chem., 74 (2002) 4370-4377. 

Manera MG, Spadavecchia J, Leone A, Quaranta F, Rella R, Dell atti D, Minunni M, Mascini M, Siciliano P (2008) Surface plasmon resonance imaging technique for nucleic acid detection. Sensors Actuators B Chem 130 82-87... 

One limitation of elecfrocatalytic detection for nucleic acid detection was its limitation to native guanine as an... 

The inclusion of the solvation correction therefore led to two important insights. The first is on the role of the deprotonation in the electrocatalytic process. The second was to modified nucleobases usefiil in sophisticated and sensitive schemes for nucleic acid detection. 

Vainrub, A. and Pettitt, B.M. Sensitive quantitative nucleic acid detection using oligonucleotide microarrays. Journal of the American Chemical Society, 2003,125 (26), p. 7798-7799. 

---