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Method for detecting nucleic acid or cells based on enzymatic cycle amplification and nano-particle reinforced SPR (surface plasmon resonance)

A technology of enzymatic cycle amplification and nano-particles, applied in biochemical equipment and methods, measuring devices, biological testing, etc., can solve the problems of long analysis time, cumbersome washing operations and separation steps, achieve good reproducibility, improve detection Sensitivity, the effect of enhancing broad applicability

Inactive Publication Date: 2016-06-08
QINGDAO UNIV OF SCI & TECH
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Problems solved by technology

Although these methods have been used in laboratory research, in practical application and clinical detection and analysis, these methods have certain limitations due to relatively long analysis time, tedious washing operations and separation steps

Method used

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  • Method for detecting nucleic acid or cells based on enzymatic cycle amplification and nano-particle reinforced SPR (surface plasmon resonance)
  • Method for detecting nucleic acid or cells based on enzymatic cycle amplification and nano-particle reinforced SPR (surface plasmon resonance)
  • Method for detecting nucleic acid or cells based on enzymatic cycle amplification and nano-particle reinforced SPR (surface plasmon resonance)

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Embodiment Construction

[0046] The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0047] 1 Experimental part

[0048] 1.1 Reagents

[0049] Carboxyl-modified magnetic microspheres (COOH-MBs, 0.1-0.5 μm, Tianjin Beisile Chromatography Technology Development Co., Ltd.); Phi29 DNA polymerase (10 U / μL, Fermentas, USA); T4 DNA ligase (5 U / μL, Thermo Fisher Corporation); E.ColiDNA Polymerase Klenow (Dalian Bao Biological Engineering Co., Ltd.); Trichloroethyl Phosphate (TCEP, 98%, AlfaAesar Company); 1–(3–Dimethylaminoaldehyde)–3– Ethyldiimine hydrochloride (EDC, Tianjin BASF Chemical Co., Ltd.); deoxynucleoside triphosphate dNTPs (Dalian Bao Biological Engineering Co., Ltd.); hydroxysuccinimide (NHS, Sigma-Aldrich Company); hydrochloric acid ( HCl, Yantai Sanhe Chemical Reagent Co., Ltd.); imidazole (Yuanyang Chemical Reagent Company); trisodium citrate (trisodiumcitrate, Tianjin Bodi Chemical Co., Ltd.); tetrachloroauric acid (HAuCl 4...

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Abstract

The invention discloses a method for detecting nucleic acid or cells based on enzymatic cycle amplification and nano-particle reinforced SPR (surface plasmon resonance). Strand displacement cycle amplification and rolling circle replication amplification are triggered by a target DNA (deoxyribonucleic acid), a double-signal amplification reaction is formed, a magnetic bead is taken as a DNA carrier and serves as a transfer station for signal amplification, a magnetic bead bio-barcode probe has a strand displacement cycle amplification reaction as a reaction unit and also increases the load capacity of a rolling circle replication product as a signal amplification carrier, a multi-stage signal amplification system is formed, high-sensitivity detection of the target DNA is realized with gold nano-particles as signal probes with an SPR technique, and the limit of detection can reach 1fM.

Description

technical field [0001] The invention relates to a method for detecting nucleic acid or cells based on enzyme cycle amplification and nanoparticle-enhanced SPR. Background technique [0002] Malignant tumor (referred to as cancer) is one of the malignant diseases that seriously threaten human life and health. The mortality rate caused by tumors in my country ranks second among all causes, and the incidence rate is on the rise. In recent years, with the continuous deepening of human research on cancer, people have gradually realized that cancer prevention is the most effective weapon against cancer. Therefore, the early detection and diagnosis of cancer is of great significance to the prevention and treatment of cancer. Tumor cells are an important indicator for diagnosing tumors. Early diagnosis and treatment of cancer can be achieved through the detection of differentiated proteins and cytokines on the surface of malignant tumor cells, and they play a key role in prognosis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/06G01N33/574G01N33/569G01N33/543G01N33/53
CPCG01N33/5308G01N33/54326G01N33/56966G01N33/574C12Q1/6804C12Q1/6844C12Q2531/119C12Q2531/125C12Q2525/301C12Q2525/205C12Q2563/143C12Q2563/149C12Q2565/518C12Q2565/628
Inventor 何鹏韩文豪乔文苹张书圣
Owner QINGDAO UNIV OF SCI & TECH
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