期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 368, 期 -, 页码 -出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2022.132179
关键词
Electrogenerated chemiluminescence; 5-Hydroxymethylcytosine; DNA nanomachine; Nanocomposite magnetic separation; Mice tissue
资金
- National Natural Science Foundation of China [22074119, 21675124]
- National Key R&D Pro-gram of China [2019YFD1002405]
- Natural Science Basic Research Program of Shaanxi [2020JM-426]
A sensitive and non-separation electrogenerated chemiluminescence (ECL) biosensing method was developed to detect 5-hydroxymethylcytosine modified double-strand DNA (5-hmC-dsDNA) using a poly (diallyl dimethyl ammonium chloride)/ferrosoferric oxide/molybdenum disulfide (PDDA/Fe3O4/MoS2) nanocomposite and DNA nanomachine. The method overcame steric resistance by fixing the negatively charged 5-hmC-dsDNA onto the positively charged and magnetic PDDA/Fe3O4/MoS2 nanocomposite, and magnetic fields separation was possible. The method showed a broader linear range and a low limit of detection, and it demonstrated potential for early clinical diagnosis of disease.
Herein, a sensitive and non-separation electrogenerated chemiluminescence (ECL) biosensing method that combines poly (diallyl dimethyl ammonium chloride)/ferrosoferric oxide/molybdenum disulfide (PDDA/Fe3O4/ MoS2) nanocomposite with DNA nanomachine was developed to determine 5-hydroxymethylcytosine modified double-strand DNA (5-hmC-dsDNA). A positively charged and magnetic PDDA/Fe3O4/MoS2 nanocomposite was proposed, which can be used to fix negatively charged 5-hmC-dsDNA to overcome the steric resistance problem and is available for magnetic fields separation. In addition, the DNA nanomachine with three elements was fabricated: a DNA walking probe for signal amplification, an ECL reagent labeled S1 DNA (Ru-S1), 5-hmC antibody (Ab) for specifically recognizing 5-hmC modification. In the presence of 5-hmC-dsDNA, sandwich type conjugates were generated to incubate PDDA/Fe3O4/MoS2 and DNA nanomachine. After magnetic separation and the DNA walking driven by enzyme cleavage, a lot of ECL reagents were released and increased the ECL signal. ECL measurement was performed in a separation-free format. The method showed a broader linear range (0.0090-0.5761 %) and a low limit of detection (0.0024 %). Furthermore, the determination of 5-hmCdsDNA in different mice tissues using the ECL biosensing method was also carried out, demonstrating that the established strategy may pave a novel avenue for the early clinical diagnosis of disease.
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