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Chemiluminescence immunoassay for cardiac troponin T by using silver nanoparticles functionalized with hemin/G-quadruplex DNAzyme on a glass chip array

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Abstract

The authors describe an array for chemiluminescence (CL) based determination of cardiac troponin T (cTnT), an important cardiovascular disease marker. The tracing tag consists of silver nanoparticles (AgNPs) loaded with guanine-rich DNA sequences and detection antibody in a high numerical ratio. The loaded AgNPs were then reacted with hemin to form a hemin/G-quadruplex DNAzyme. A disposable immunosensor array was fabricated by immobilizing capture antibody on corresponding sensing sites on a glass chip. Once a sandwich immunocomplex is formed on the array, the tracing tag catalyzes the CL reaction of the luminol-p-iodophenol and H2O2 system to produce a CL signal, which is collected by a CCD camera. An intuitive CL image is obtained containing all of the spots on the array. Under optimal conditions, the method shows a wide linear range over 4 orders of magnitude (from 0.003 to 270 ng·L−1), a detection limit down to 84 fg·L−1, and a throughput as high as 44 tests·h−1. The results obtained with serum samples are in acceptable agreement with reference values. The AgNP-based tracing tag as well as the immunoassay method shows a promising potential for point-of-care testing for the early clinical diagnosis of cardiovascular disease.

Schematic presentation of silver nanoparticles (AgNPs) functionalized with hemin/G-quadruplex DNAzyme for highly sensitive chemiluminescence (CL) immunoassay of cardiac troponin T (cTnT) on a glass chip array.

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Acknowledgements

This work is supported by the National Natural Science Foundation of China (81421005, 21505160), Natural Science Foundation of Jiangsu Province (BK20150690), the Fundamental Research Funds for the Central Universities (2015PY009) and State Key Laboratory Analytical Chemistry for Life Science (SKLACLS1513).

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Author notes

  1. Chen Zong and Duo Duo Zhang contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, 210009, China

    Chen Zong, Duoduo Zhang, Hua Yang & Ping Li

  2. College of Traditional Chinese Medicine, Guangdong Pharmaceutical University, Guangzhou, 510006, China

    Duoduo Zhang, Shumei Wang & Ping Li

  3. The First Affiliated Hospital with Nanjing Medical University, Nanjing, 210029, China

    Ming Chu

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  1. Chen Zong
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  2. Duoduo Zhang
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Correspondence to Ping Li.

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Zong, C., Zhang, D., Yang, H. et al. Chemiluminescence immunoassay for cardiac troponin T by using silver nanoparticles functionalized with hemin/G-quadruplex DNAzyme on a glass chip array. Microchim Acta 184, 3197–3204 (2017). https://doi.org/10.1007/s00604-017-2331-z

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