Recombinase polymerase amplification combined with a magnetic nanoparticle-based immunoassay for fluorometric determination of troponin T

Target‐Dependent Protection of DNA Aptamers against Nucleolytic Digestion Enables Signal‐On Biosensing with Toehold‐Mediated Rolling Circle Amplification

“…[2] More recently, ITA methods have been used for detection of non-nucleic acid species, with many assays employing creative methods to allow molecular recognition elements to modulate the degree of ITA. These include: nucleic acid modified antibodies, [3][4][5] antibody-directed proximity ligation assays, [6][7][8] and various methods to couple functional nucleic acids (FNAs) such as aptamers [9][10][11][12][13] and DNAzymes to ITA methods. [14][15][16][17] Common drawbacks of these strategies include the requirement for specially modified antibodies and separation steps for immuno-coupled ITA methods, optical or electrochemical labelling of nucleic acids, multiple primers or processing enzymes for most of the FNA-coupled ITA methods, or the need for elevated reaction temperatures.…”

The CRISPR–Cas toolbox for analytical and diagnostic assay development

“…The general strategy is to quantitatively convert the non-nucleic-acid targets into predesigned DNA or RNA intermediates that can then be recognized and detected by CRISPR-Cas12 or Cas13 systems. Mechanistically, this quantitative conversion can be achieved through a classic immunoassay format, [154][155][156] or the use of functional nucleic acids, 151,153 or in a homogenous solution through proximity binding. 157 Ever since the development of immune-PCR, the combination between immunoassay and nucleic acid amplification has been widely adapted for the ultrasensitive detection of proteins and small molecules.…”

“…157 Ever since the development of immune-PCR, the combination between immunoassay and nucleic acid amplification has been widely adapted for the ultrasensitive detection of proteins and small molecules. [154][155][156] A similar principle has also been adapted to CRISPR-Cas systems for the amplified detection of non-nucleic-acid targets. For instance, Chen et al developed a CRISPR-Cas13a signal amplification linked immunosorbent assay (CLISA) by employing Cas13a as a signal amplifier to a classic immunoassay (Fig.…”

Nucleic Acid Amplification Techniques in Immunoassay: An Integrated Approach with Hybrid Performance

“…In recent years, recombinase polymerase amplification (RPA) has gradually been used in immunoassays. As shown in Figure , Ivanov et al designed an RPA immunosensor with magnetic nanoparticles (MNPs) which conjugate with primary antibodies. First, the secondary antibodies are combined with reporter DNA via biotin–streptavidin interaction.…”