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A triple-amplification colorimetric assay for antibiotics based on magnetic aptamer-enzyme co-immobilized platinum nanoprobes and exonuclease-assisted target recycling.
Analyst. 2015 Nov 21; 140(22):7663-71.A

Abstract

Herein, an ultrasensitive and selective colorimetric assay for antibiotics, using chloramphenicol (CAP) as the model analyte, was developed based on magnetic aptamer-HRP-platinum composite probes and exonuclease-assisted target recycling. The composite probes were prepared through immunoreactions between the double stranded DNA antibody (anti-DNA) labeled on core-shell Fe3O4@Au nanoparticles (AuMNP-anti-DNA) as the capture probe, and the double stranded aptamer (aptamer hybrid with its complementary oligonucleotides) labeled on Pt@HRP nanoparticles as the nanotracer (ds-Apt-HRP-PtNPs). When the CAP samples were incubated with the probes for 30 min at room temperature, they could be captured by the aptamer to form a nanotracer-CAP complex, which was then released into the supernatant after magnetic separation. This is because the anti-DNA on the capture probes cannot recognize the single strand aptamer-CAP complex. The exonuclease I (Exo I) added into the supernatant can further digest the aptamer-CAP from the 3'-end of the aptamer and the CAP in the aptamer-CAP complex can be released again, which can further participate in a new cycling process to react with the probes. Pt and HRP in the nanotracer could both catalyze and dual amplify the absorbance at 650 nm ascribed to the 3,3',5,5'-tetramethylbenzidine (TMB)-H2O2 system. Moreover, Exo I can assist the target recycling, which can further amplify the signal. Thus, the triple amplified signal can be quantified by ultraviolet-visible spectroscopy. The experimental results showed that the CAP detection possessed a linear range of 0.001-10 ng mL(-1) and a detection limit of 0.0003 ng mL(-1) (S/N = 3). The assay was successfully employed to detect CAP in milk, which is much more facile, time saving, and sensitive than the commercial ELISA kits.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, China.

Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, China.

Key Laboratory of Asymmetric Synthesis and Chirotechnology of Sichuan Province, Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu, 610041, China.

Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, China.

Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, China.

Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, China.

Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, China.

Faculty of Food Science and Engineering, Nanjing University of Finance and Economics, Nanjing, 210000, China.

MeSH

AnimalsAnti-Bacterial AgentsAptamers, NucleotideBenzidinesBiosensing TechniquesChloramphenicolColorimetryExodeoxyribonucleasesGoldHydrogen PeroxideLimit of DetectionMagnetite NanoparticlesMilkPlatinum

Pub Type(s)

Evaluation Study

Journal Article

Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

26442572

Citation

Miao, Yangbao, et al. "A Triple-amplification Colorimetric Assay for Antibiotics Based On Magnetic Aptamer-enzyme Co-immobilized Platinum Nanoprobes and Exonuclease-assisted Target Recycling." The Analyst, vol. 140, no. 22, 2015, pp. 7663-71.

Miao Y, Gan N, Ren HX, et al. A triple-amplification colorimetric assay for antibiotics based on magnetic aptamer-enzyme co-immobilized platinum nanoprobes and exonuclease-assisted target recycling. Analyst. 2015;140(22):7663-71.

Miao, Y., Gan, N., Ren, H. X., Li, T., Cao, Y., Hu, F., Yan, Z., & Chen, Y. (2015). A triple-amplification colorimetric assay for antibiotics based on magnetic aptamer-enzyme co-immobilized platinum nanoprobes and exonuclease-assisted target recycling. The Analyst, 140(22), 7663-71. https://doi.org/10.1039/c5an01142f

Miao Y, et al. A Triple-amplification Colorimetric Assay for Antibiotics Based On Magnetic Aptamer-enzyme Co-immobilized Platinum Nanoprobes and Exonuclease-assisted Target Recycling. Analyst. 2015 Nov 21;140(22):7663-71. PubMed PMID: 26442572.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - A triple-amplification colorimetric assay for antibiotics based on magnetic aptamer-enzyme co-immobilized platinum nanoprobes and exonuclease-assisted target recycling. AU - Miao,Yangbao, AU - Gan,Ning, AU - Ren,Hong-Xia, AU - Li,Tianhua, AU - Cao,Yuting, AU - Hu,Futao, AU - Yan,Zhongdan, AU - Chen,Yinji, PY - 2015/10/8/entrez PY - 2015/10/8/pubmed PY - 2016/7/23/medline SP - 7663 EP - 71 JF - The Analyst JO - Analyst VL - 140 IS - 22 N2 - Herein, an ultrasensitive and selective colorimetric assay for antibiotics, using chloramphenicol (CAP) as the model analyte, was developed based on magnetic aptamer-HRP-platinum composite probes and exonuclease-assisted target recycling. The composite probes were prepared through immunoreactions between the double stranded DNA antibody (anti-DNA) labeled on core-shell Fe3O4@Au nanoparticles (AuMNP-anti-DNA) as the capture probe, and the double stranded aptamer (aptamer hybrid with its complementary oligonucleotides) labeled on Pt@HRP nanoparticles as the nanotracer (ds-Apt-HRP-PtNPs). When the CAP samples were incubated with the probes for 30 min at room temperature, they could be captured by the aptamer to form a nanotracer-CAP complex, which was then released into the supernatant after magnetic separation. This is because the anti-DNA on the capture probes cannot recognize the single strand aptamer-CAP complex. The exonuclease I (Exo I) added into the supernatant can further digest the aptamer-CAP from the 3'-end of the aptamer and the CAP in the aptamer-CAP complex can be released again, which can further participate in a new cycling process to react with the probes. Pt and HRP in the nanotracer could both catalyze and dual amplify the absorbance at 650 nm ascribed to the 3,3',5,5'-tetramethylbenzidine (TMB)-H2O2 system. Moreover, Exo I can assist the target recycling, which can further amplify the signal. Thus, the triple amplified signal can be quantified by ultraviolet-visible spectroscopy. The experimental results showed that the CAP detection possessed a linear range of 0.001-10 ng mL(-1) and a detection limit of 0.0003 ng mL(-1) (S/N = 3). The assay was successfully employed to detect CAP in milk, which is much more facile, time saving, and sensitive than the commercial ELISA kits. SN - 1364-5528 UR - https://www.unboundmedicine.com/medline/citation/26442572/A_triple_amplification_colorimetric_assay_for_antibiotics_based_on_magnetic_aptamer_enzyme_co_immobilized_platinum_nanoprobes_and_exonuclease_assisted_target_recycling_ DB - PRIME DP - Unbound Medicine ER -