phthalic-acid-ester-binding-dna-aptamer-selection-characterization

Phthalic Acid Ester-Binding DNA Aptamer Selection, Characterization, and Application to an Electrochemical Aptasensor

Phthalic acid esters (PAEs) areone of the major groups of persistent organic pollutants. The group-specific detection of PAEs is highly desired due to the ...

Traditional methods for identifying food-borne pathogens are time-consuming and laborious, so it is necessary to develop innovative methods for the rapid identification of food-borne pathogens. Here, we report the development of silicon-based optical thin-film biosensor chips for sensitive detection of 11 food-borne pathogens. Briefly, aldehyde-labeled probes were arrayed and covalently attached to a hydrazine-derivatized chip surface, and then, biotinylated polymerase chain reaction (PCR) amplicons were hybridized with the probes. After washing and brief incubation with an antibiotin immunoglobulin G-horseradish peroxidase conjugate and a precipitable horseradish peroxidase substrate, biotinylated chains bound to the probes were visualized as a color change on the chip surface (gold to blue/purple). Highly sensitive and accurate examination of PCR fragment targets can be completed within 30 min. This assay is extremely robust, sensitive, specific, and economical and can be adapted to different throughputs. Thus, a rapid, sensitive, and reliable technique for detecting 11 food-borne pathogens was successfully developed.

Rapid and reliable detection of 11 food-borne pathogens using thin-film biosensor chips.

Traditionelle Methoden zur Identifizierung von lebensmittelbedingten Krankheitserregern sind zeitaufw&auml;ndig und m&uuml;hsam, so ist es erforderlich, innovative Methoden zur schnellen Identifizierung von Lebensmittel &uuml;bertragbaren Krankheitserregern zu entwickeln. Hier berichten wir &uuml;ber die Entwicklung der Biosensor f&uuml;r optische D&uuml;nnschicht-Silizium-basierten Chips f&uuml;r empfindliche Nachweis von 11 lebensmittelbedingten Krankheitserregern. Kurz gesagt, Aldehyd-markierten Sonden gekleidet kovalent gebunden zu einer Hydrazin-derivatized Chip-Oberfl&auml;che und wurden dann, biotinylierten Polymerase-Kettenreaktion (PCR) Amplifikate wurden mit den Sonden hybridisiert. Nach dem Waschen und kurze Inkubation mit ein antibiotin Immunglobulin G-Meerrettich-Peroxidase-Konjugat und einem Ausf&auml;llbares Meerrettich Peroxydase Substrat, biotinylierten Ketten gebunden um die Sonden wurden als eine Farb&auml;nderung auf der Chip-Oberfl&auml;che (Gold, blau/violett) visualisiert. Hochempfindliche und genaue Pr&uuml;fung der PCR-Fragment-Ziele kann innerhalb von 30 Minuten abgeschlossen werden. Diese Probe ist extrem robust, sensibel, spezifische und wirtschaftlich und kann an verschiedene Durchs&auml;tze angepasst werden. Dazu wurde eine schnelle, vertrauliche und verl&auml;ssliche Technik zum Erkennen von 11 lebensmittelbedingten Krankheitserregern erfolgreich entwickelt.

Schnelle und zuverl&auml;ssige Erkennung von 11 lebensmittelbedingten Krankheitserregern mit D&uuml;nnschicht-Biosensor-Chips.

To observe the analgesic effect of CQM on photochemically-induced prosopalgia model rats, and discuss its impact on the exciting amino acid neurotransmitter-glutamate (Glu).

[Analgesic effect of CQM on prosopalgia model rats and its impact on exciting amino acid neurotransmitters].

A wide range of pathologies have been targeted with bimodular aptamers that contain both G-quadruplex (G4) and duplex motifs, while the structures and functions are poorly understood. G4-selective fluorescent dyes have served as facile tools to probe G4s, but not for bimodular aptamers, yet. Here, taking the 29-mer thrombin binding aptamer (TBA29) as an example, we demonstrated that 3,6-dimethyl-2-(4-dimethylaminophenyl)-benzothiazolium (ThT) was the most effective dye compared to NMM and PPIX in recognizing TBA29. Binding studies indicate that ThT recognized TBA29 via distinct buffer-dependent mechanisms. Specifically, ThT induced the formation of a bimolecular parallel G4 in cation-deficient buffer, showing 341-fold fluorescent enhancement. The competitive binding of thrombin disrupted the complex, leading to the monotonic fluorescence decrease. A similar mechanism was previously reported for the interaction between ThT and the 15-mer thrombin binding aptamer (TBA15). However, TBA29 bound with ThT in a more favorable state than TBA15, showing hyperchromic effects and two times stronger fluorescence enhancement. Differently, ThT bound with antiparallel TBA29/TBA15 in an intercalating/groove binding mode in 100 mM KCl, generating 181/28-fold fluorescence enhancement, respectively. These results revealed that ThT recognized both parallel and antiparallel G4s of TBA29 more efficiently than it recognized TBA15. The duplex structure of TBA29 may play an important role in its interaction with ThT. Our study broadens the application of ThT in screening G4 to bimodular aptamers and provides some insights into the structures of TBA29, along with the interaction between ThT and TBA29. Our study also is useful for the development of structure-switching-based biosensors using bimodular aptamers. Graphical abstract The buffer-dependent binding mechanisms of ThT with TBA29, and the competitive (top)/noncompetitive (bottom) binding of thrombin with TBA29-ThT complex.

Article	Year
Rapid and reliable detection of 11 food-borne pathogens using thin-film biosensor chips. Applied microbiology and biotechnology\| PubMed ID: 20091028	2010
[Analgesic effect of CQM on prosopalgia model rats and its impact on exciting amino acid neurotransmitters]. Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica\| PubMed ID: 24490572	2013
Thioflavin T as a fluorescence light-up probe for both parallel and antiparallel G-quadruplexes of 29-mer thrombin binding aptamer. Analytical and bioanalytical chemistry\| PubMed ID: 27590320	2016

Shi Xu

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