High-efficient separation of deoxyribonucleic acid from pathogenic bacteria by hedgehog-inspired magnetic nanoparticles microextraction.

Autor: Shi YJ; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital / Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300134, China., Che YN; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital / Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300134, China., Zhao YM; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital / Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300134, China., Ran RX; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China., Zhao YQ; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital / Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300134, China., Yu SS; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital / Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300134, China., Chen MY; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital / Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300134, China., Dong LY; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China. Electronic address: donglinyi@tmu.edu.cn., Zhao ZY; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital / Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300134, China. Electronic address: zhaozhenyu0858@163.com., Wang XH; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China. Electronic address: xianhua.w@163.com.
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2024 Jun 07; Vol. 1724, pp. 464923. Date of Electronic Publication: 2024 Apr 21.
DOI: 10.1016/j.chroma.2024.464923
Abstrakt: Efficient separation of deoxyribonucleic acid (DNA) through magnetic nanoparticles (MN) is a widely used biotechnology. Hedgehog-inspired MNs (HMN) possess a high-surface-area due to the distinct burr-like structure of hedgehog, but there is no report about the usage of HMN for DNA extraction. Herein, to improve the selection of MN and illustrate the performance of HMN for DNA separation, HMN and silica-coated Fe 3 O 4 nanoparticles (Fe 3 O 4 @SiO 2 ) were fabricated and compared for the high-efficient separation of pathogenic bacteria of DNA. Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) are typical Gram-negative and Gram-positive bacteria and are selected as model pathogenic bacteria. To enhance the extraction efficiency of two kinds of MNs, various parameters, including pretreatment, lysis, binding and elution conditions, have been optimized in detail. In most separation experiments, the DNA yield of HMN was higher than that of Fe 3 O 4 @SiO 2 . Therefore, a HMN-based magnetic solid-phase microextraction (MSPE) and quantitative real-time PCR (qPCR) were integrated and used to detect pathogenic bacteria in real samples. Interestingly, the HMN-based MSPE combined qPCR strategy exhibited high sensitivity with a limit of detection of 2.0 × 10 1 CFU mL -1 for E. coli and 4.0 × 10 1 CFU mL -1 for S. aureus in orange juice, and 2.8 × 10 2 CFU mL -1 for E. coli and 1.1 × 10 2 CFU mL -1 for S. aureus in milk, respectively. The performance of the proposed strategy was significantly better than that of commercial kit. This work could prove that the novel HMN could be applicable for the efficient separation of DNA from complex biological samples.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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