Recent advances in fluorescence resonance energy transfer-based probes in nucleic acid diagnosis

Abstract

Nucleic acid diagnosis is a method that diagnoses human conditions and diseases by directly exploring the existing states or defects of nucleic acids using theoretical and technical approaches from molecular biology.

Organomercury Nucleic Acids: Past, Present and Future

“…Various methods for the detection of single nucleotide polymorphisms (SNPs) rely on differences in the hybridization affinities of oligonucleotide probes for the target sequence. [99][100][101][102] Unfortunately, canonical Watson-Crick base pairing is less than ideal for this approach. Although the matched base pair is usually much more stable than any of the mispairs, stabilities of the latter do not differ sufficiently from each other to allow reliable identification of the polymorphic nucleobase (Figure 4A).…”

“…Various methods for the detection of single nucleotide polymorphisms (SNPs) rely on differences in the hybridization affinities of oligonucleotide probes for the target sequence [99–102] . Unfortunately, canonical Watson‐Crick base pairing is less than ideal for this approach.…”

Construction of Aptamer-Based Molecular Beacons with Varied Blocked Structures and Targeted Detection of Thrombin

“…The molecular beacon (MB) itself can form a closed hairpin structure, including the loop portion and the stem portion; the loop portion contains the antisense recognition domain or the aptamer sequence of the target. − In a conventional MB, a fluorophore/quencher pair is introduced and labeled on either end. MB relies on a “closed” configuration by the stem part in the absence of a target, which causes the quenching of the fluorophore by the fluorescence resonance energy transfer (FRET). − When the MB encounters a target object, the hairpin structure spontaneously opens with the disruption of the stem and results in a fluorescence restoration. − The MB shows many advantages over other oligonucleotide probes, including strong specificity and high sensitivity. , It has been widely applied in chemistry, biology, medicine, and other fields, especially the detection of protein and polypeptide, mRNA monitoring, real-time imaging of living cells, and so on.…”

Rapid and Wash-Free Time-Gated FRET Histamine Assays Using Antibodies and Aptamers

“…Förster resonance energy transfer (FRET) is a strong donor–acceptor distance-dependent energy transfer mechanism that has been widely used for the quantification of biomolecular distances and concentrations via fluorescence biosensing. − FRET-based immunoassays are usually sandwich assays (binding of donor–antibody and acceptor–antibody to the same target results in FRET) or competitive assays (free target competes with donor or acceptor labeled target that is bound to an acceptor- or donor-labeled antibody) and have the advantage of simpler and quicker wash-free procedures (the binding-related FRET signal is different from other nonspecific signals) and more precise ratiometric detection (if both donor and acceptor are fluorescent). ,, When lanthanide-based donors with very long (up to milliseconds) excited-state lifetimes are used in FRET, the use of time-gated (TG) detection can efficiently reduce sample autofluorescence and other interfering signals, resulting in highly sensitive homogeneous assay formats. ,− Using terbium (Tb) donors and dye or quantum dot acceptors, we have developed noncompetitive TG-FRET assays against various protein or nucleic acid biomarkers. ,− However, implementing TG-FRET into specific, sensitive, rapid, and wash-free competitive assays for small molecules, such as HA, remains to be demonstrated. For an efficient translation into different sensing concepts, it would also be highly desirable if such a homogeneous HA assay could be generically applied to different molecular recognition molecules, such as antibodies and aptamers.…”