Evaluation of Relative Aptamer Binding to<i>Campylobacter jejuni</i>Bacteria Using Affinity Probe Capillary Electrophoresis

Stratis‐Cullum, Dimitra N.; McMasters, Sun; Pellegrino, Paul M.

doi:10.1080/00032710903137376

Cited by 16 publications

(5 citation statements)

References 29 publications

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“…Our group has demonstrated that aptamers it has developed against Campylobacter jejuni and Listeria monocytogenes can be used in QD or enzymelinked fluorescence aptamer-paramagnetic bead (MB) sandwich assays to achieve detection limits below 10 colony forming units (cfu) or cells per ml [13,15]. Our aptamers and ultralow detection limits even in various foods or culture enrichment broths have been validated by other researchers [11,[16][17][18] and independent laboratories [13] and led to a recent second place finish among 49 competitors in the U.S. FDA's inaugural Food Safety Testing Technology Challenge [30][31][32]. While, our aptamers have proven useful for improving detection limits in traditional LF test strip formats with QDs [4] or tube-based MB fluorescent sandwich assays [13,15], they have thus far only been applied to single target tests.…”

Section: Introductionmentioning

confidence: 86%

“…Therefore, our group and many others have been investigating application of aptamers for food safety assays [4,[12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29], because food safety testing is an area of "zero tolerance, " i.e., no detectable foodborne pathogens are allowed in tested foods or enrichment broth cultures derived from tested foods. Thus, ultrasensitivity is desired in food safety assays to reduce culture enrichment times enabling faster clearance and sale of refrigerated or frozen food products which saves money for food producers and increases profitability.…”

Section: Introductionmentioning

confidence: 99%

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Preliminary Development of a Magnetically Assisted Test Strip (MATS) Cartridge and Fluorescent DNA Aptamer-Magnetic Bead Quantum Dot Sandwich Assays for Multiplexed Food Safety Applications

Bruno¹,

Phillips²

2016

J Biomed Eng Med Devic

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Preliminary development of a simple mesofluidic multi-channel plastic cartridge with underlying external magnet to drag DNA aptamer-coated paramagnetic beads through fluids in the channels while developing a sandwich assay with quantum dot-conjugated reporter aptamers is described. This approach is superior to traditional lateral flow test strips in several ways including: 1) the ability to control the speed of lateral flow in the channel versus conventional nitrocellulose analytical membranes with fixed wicking times. 2) The use of aptamers for potentially greater affinity and consistency from batch-to-batch versus comparable antibodies. 3) Superior fluorescence efficiency and intensity provided by quantum dots versus conventional fluorescent dyes and 4) the ability to multiplex based on the various colored emissions of different sized quantum dots when excited with a single ultraviolet source. Development of the system from concept to prototype is described along with illustration of sensitive system performance for several food safety-related targets. The system is also clearly adaptable to rapid multiplex detection and sensitive quantitation of clinical biomarkers, drugs, environmental, veterinary or other target analytes.

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Section: Introductionmentioning

confidence: 86%

Section: Introductionmentioning

confidence: 99%

Preliminary Development of a Magnetically Assisted Test Strip (MATS) Cartridge and Fluorescent DNA Aptamer-Magnetic Bead Quantum Dot Sandwich Assays for Multiplexed Food Safety Applications

Bruno¹,

Phillips²

2016

J Biomed Eng Med Devic

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“…For example, G‐protein‐BODIPY FL GTPγS, hairpin‐structured DNA‐peptide nucleic acid, concanavalin A‐monosaccharides, dorzolamide‐human carbonicanhydrase II, and tubulin‐anti‐mitotic compounds pairs were examined . Other examples of the aptamer‐based ACE applications are summarized in Table .…”

Section: Aptamer‐based Affinity Cementioning

confidence: 99%

Capillary electrophoresis–based immunoassay and aptamer assay: A review

2020

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Capillary electrophoresis-based immunoassay and aptamer assay: A reviewOver the last two decades, the group of techniques called affinity probe CE has been widely used for the detection and the determination of several types of biomolecules with high sensitivity. These techniques combine the low sample consumption and high separation power of CE with the selectivity of the probe to the target molecule. The assays can be defined according to the type of probe used: CE immunoassays, with an antibody as the probe, or aptamer-based CE, with an aptamer as the probe. Immunoassays are generally divided into homogeneous and heterogeneous groups, and homogeneous variant can be further performed in competitive or noncompetitive formats. Interacting partners are free in solution at homogeneous assay, as opposed to heterogeneous analyses, where one of them is immobilized onto a solid support. Highly sensitive fluorescence, chemiluminescence or electrochemical detections were typically used in this type of study. The use of the aptamers as probes has several advantages over antibodies such as shorter generation time, higher thermal stability, lower price, and lower variability. The aptamer-based CE technique was in practice utilized for the determination of proteins in biological fluids and environmentally or clinically important small molecules. Both techniques were also transferred to microchip. This review is focused on theoretical principles of these techniques and a summary of their applications in research.Abbreviations: Ag, antigen; CEIA, CE immunoassay; MCE, microchip CE; SELEX, systematic evolution of ligands by exponential enrichment and they are produced by the immune system of living organisms. Ag that start the immunological response are also called immunogens. These Ab-Ag complexes are stabilized by dipole-dipole, van der Waals or hydrophobic interactions and hydrogen bonds [5,6].The most often used immunoassays are ELISA with Ab or Ag immobilized onto solid support (plates, glass fibers, or plastic tubes), immunofluorescence assay, fluorescence polarization immunoassay, luminescence immunoassay, microparticle enzyme immunoassay, and biosensors. The major disadvantages of immunoassays are the tedious process, time-consuming reactions, and problems with reproducibility and nonspecific interactions [6].CE is a powerful analytical method for the separation of a wide range of molecules. The coupling of CE and immunoassay in the new CE immunoassay (CEIA) technique provided both separation power and specificity of interactions. Other advantages are the possibility of multianalyte analyses, automation, low sample consumption, high speed, flexibility, a lower amount of false-positive results, and also the possibility of connection with highly sensitive detectors [5].In CEIA, the Fab fragments of Ab containing Ag-specific regions are more often used than the whole monoclonal and Color online: See article online to view Figs. 4 and 7 in color.

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“…These aptamers showed high affinity and specificity against the target with relative high dissociation constant of 292.8 ± 53.1 nM (Jaykus 2010;Dwivedi et al 2010b). Furthermore, Stratis-Cullum and others performed a CE-SELEX to evaluate the aptamer specific for dead C. jejuni with qualitative capillary electrophoresis immunoassay and the obtained LOD was 6.3 × 10 6 cells/mL (Stratis-Cullum et al 2009). Aptamer-linked QDs have been developed and used in a sandwich assay in conjugation with magnetic beads for the detection of C. jejuni at levels of between 2.5 and 10-50 cfu/ mL in buffer and food matrices, respectively (Bruno et al 2009).…”

Section: Campylobacter Jejunimentioning

confidence: 99%

An Update on Aptamer-Based Multiplex System Approaches for the Detection of Common Foodborne Pathogens

et al. 2017

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Foodborne ailments constitute a public health challenge and pose an incredible economic burden in healthcare system around the globe. This dilemma has urged authorities and other entities working in field of food quality control and supply chain to play a pivotal role in ensuring food safety. Analytical strategies have been developed using numerous systematic evolution of ligands by exponential enrichment (SELEX) methods to assure food safety. High-affinity and high-sensitivity ssDNA and RNA aptamers against pathogens have emerged as a novel strategy, as compared to the more resource-demanding and complicated biochemical test-based approaches. Thus, this review aims to focus on some methods used in the selection of specific bare, modified, and conjugated aptamers and on the further analysis of selected aptamers using flow cytometer or post-SELEX modifications for enhanced detection of frequently diagnosed foodborne bacteria such as Bacillus sp., Campylobacter jejuni, Escherichia sp., Salmonella sp., Staphylococcus aureus, Shigella sp., Listeria monocytogenes, and Streptococcus pyogenes and/or targeting their cell components towards attaining fast, sensitive, and selective methods for the detection of pathogens in food(s) or other sources.

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Evaluation of Relative Aptamer Binding toCampylobacter jejuniBacteria Using Affinity Probe Capillary Electrophoresis

Cited by 16 publications

References 29 publications

Preliminary Development of a Magnetically Assisted Test Strip (MATS) Cartridge and Fluorescent DNA Aptamer-Magnetic Bead Quantum Dot Sandwich Assays for Multiplexed Food Safety Applications

Preliminary Development of a Magnetically Assisted Test Strip (MATS) Cartridge and Fluorescent DNA Aptamer-Magnetic Bead Quantum Dot Sandwich Assays for Multiplexed Food Safety Applications

Capillary electrophoresis–based immunoassay and aptamer assay: A review

An Update on Aptamer-Based Multiplex System Approaches for the Detection of Common Foodborne Pathogens

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