Bioanalytical applications of aptamer and molecular-beacon probes in fluorescence-affinity assays

Goulko, Alevtina A.; Li, Feng; Le, X. Chris

doi:10.1016/j.trac.2009.03.014

Cited by 55 publications

(34 citation statements)

References 81 publications

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“…Aptamers have been applied in protein detection [17], [18], diseases diagnosis [19], [20], cell imaging and tracking [21], [22] ect. owing to the advantages of exceptional stability, easy manipulation and reproducibility, non-toxic and diagnostic potential.…”

Section: Introductionmentioning

confidence: 99%

A Visual Dual-Aptamer Logic Gate for Sensitive Discrimination of Prion Diseases-Associated Isoform with Reusable Magnetic Microparticles and Fluorescence Quantum Dots

Xiao

Chen

et al. 2013

PLoS ONE

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Molecular logic gates, which have attracted increasing research interest and are crucial for the development of molecular-scale computers, simplify the results of measurements and detections, leaving the diagnosis of disease either “yes” or “no”. Prion diseases are a group of fatal neurodegenerative disorders that happen in human and animals. The main problem with a diagnosis of prion diseases is how to sensitively and selectively discriminate and detection of the minute amount of PrPRes in biological samples. Our previous work had demonstrated that dual-aptamer strategy could achieve highly sensitive and selective discrimination and detection of prion protein (cellular prion protein, PrPC, and the diseases associated isoform, PrPRes) in serum and brain. Inspired by the advantages of molecular logic gate, we further conceived a new concept for dual-aptamer logic gate that responds to two chemical input signals (PrPC or PrPRes and Gdn-HCl) and generates a change in fluorescence intensity as the output signal. It was found that PrPRes performs the “OR” logic operation while PrPC performs “XOR” logic operation when they get through the gate consisted of aptamer modified reusable magnetic microparticles (MMPs-Apt1) and quantum dots (QDs-Apt2). The dual-aptamer logic gate simplifies the discrimination results of PrPRes, leaving the detection of PrPRes either “yes” or “no”. The development of OR logic gate based on dual-aptamer strategy and two chemical input signals (PrPRes and Gdn-HCl) is an important step toward the design of prion diseases diagnosis and therapy systems.

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Section: Introductionmentioning

confidence: 99%

A Visual Dual-Aptamer Logic Gate for Sensitive Discrimination of Prion Diseases-Associated Isoform with Reusable Magnetic Microparticles and Fluorescence Quantum Dots

Xiao

Chen

et al. 2013

PLoS ONE

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“…13,16 To further increase the overall signal, amplification has been carried out through encapsulation of thousands of HRP 17,18 , quantum dots 19 or organic dyes 20,21 in various types of nanoparticles; or via nucleic acid-based enzymatic reactions. 22,23 Although very low detection limits could be achieved, 23 the complicated reaction designs and inadequate robustness hinder the more general applications in fields like point-of-care diagnosis and on-site biosafety monitoring.…”

Section: Introductionmentioning

confidence: 99%

Detection of Femtomolar Proteins by Nonfluorescent ZnS Nanocrystal Clusters

et al. 2012

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Cation exchange (CX) in the non-fluorescent ZnS nanocrystal clusters (NCCs) was employed to detect trace biomolecules with immunoassays. The NCCs were porous, and allowed fast cation exchange reaction to release an ultra-large number of Zn2+ from each cluster that turned on the Zn-responsive dyes for fluorescence detection. The ZnS NCCs were highly stable in biological buffers and more biocompatible than quantum dots. Zn2+ release efficiency and target binding by NCCs with average diameters of 44 nm, 86 nm, and 144 nm were investigated. The smallest NCCs exhibited the highest CX efficiency because of its larger surface area and bigger pores inside the cluster structure, and 71.0% of the enclosed Zn2+ were freed by CX with 2-minute microwave irradiation. They also experienced the least space hindrance and the fastest rate when binding to target molecules immobilized on surface. When the 44-nm NCCs were used to detect IgE in a sandwich assay, the limit of detection (LOD) was 5 pg/mL (33 fM), 1,000 times better than that of ELISA. Our results well demonstrate that CX in the ZnS NCCs is superior to the conventional signaling strategies in its high amplification efficiency, robustness, and biocompatibility.

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“…Because the loop is much longer than the stem, the perfect match between the loop and the target sequence determines the conformational change of the probe and disassociation of the stem. Well-designed molecular beacons with 1 mismatched base in their loops with the target sequences would not result in disassociation of the stem [8,9]. Accordingly, molecular beacons offer many advantages over ordinary DNA probes in detecting point mutations, such as higher sensitivity, specificity, and reusability (Figure 1).…”

Section: Introductionmentioning

confidence: 99%

A novel asymmetric-loop molecular beacon-based two-phase hybridization assay for accurate and high-throughput detection of multiple drug resistance-conferring point mutations in Mycobacterium tuberculosis

Chen

Xie

et al. 2012

Med Sci Monit

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SummaryThe accurate and high-throughput detection of drug resistance-related multiple point mutations remains a challenge. Although the combination of molecular beacons with bio-immobilization technology, such as microarray, is promising, its application is difficult due to the ineffective immobilization of molecular beacons on the chip surface. Here, we propose a novel asymmetric-loop molecular beacon in which the loop consists of 2 parts. One is complementary to a target, while the other is complementary to an oligonucleotide probe immobilized on the chip surface. With this novel probe, a two-phase hybridization assay can be used for simultaneously detecting multiple point mutations. This assay will have advantages, such as easy probe availability, multiplex detection, low background, and high-efficiency hybridization, and may provide a new avenue for the immobilization of molecular beacons and high-throughput detection of point mutations.

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Bioanalytical applications of aptamer and molecular-beacon probes in fluorescence-affinity assays

Cited by 55 publications

References 81 publications

A Visual Dual-Aptamer Logic Gate for Sensitive Discrimination of Prion Diseases-Associated Isoform with Reusable Magnetic Microparticles and Fluorescence Quantum Dots

A Visual Dual-Aptamer Logic Gate for Sensitive Discrimination of Prion Diseases-Associated Isoform with Reusable Magnetic Microparticles and Fluorescence Quantum Dots

Detection of Femtomolar Proteins by Nonfluorescent ZnS Nanocrystal Clusters

A novel asymmetric-loop molecular beacon-based two-phase hybridization assay for accurate and high-throughput detection of multiple drug resistance-conferring point mutations in Mycobacterium tuberculosis

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Bioanalytical applications of aptamer and molecular-beacon probes in fluorescence-affinity assays

Cited by 55 publications

References 81 publications

A Visual Dual-Aptamer Logic Gate for Sensitive Discrimination of Prion Diseases-Associated Isoform with Reusable Magnetic Microparticles and Fluorescence Quantum Dots

A Visual Dual-Aptamer Logic Gate for Sensitive Discrimination of Prion Diseases-Associated Isoform with Reusable Magnetic Microparticles and Fluorescence Quantum Dots

Detection of Femtomolar Proteins by Nonfluorescent ZnS Nanocrystal Clusters

A novel asymmetric-loop molecular beacon-based two-phase hybridization assay for accurate and ­high-throughput detection of multiple drug resistance-conferring point mutations in Mycobacterium tuberculosis

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Product

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A novel asymmetric-loop molecular beacon-based two-phase hybridization assay for accurate and high-throughput detection of multiple drug resistance-conferring point mutations in Mycobacterium tuberculosis