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Silicon nanowire chip simultaneously detecting miRNAs and protein markers and detection method and application of silicon nanowire chip

A silicon nanowire and protein labeling technology, which is applied in nanotechnology for sensing, nanotechnology for information processing, nanotechnology, etc., can solve cumbersome step processing, sensitivity and specificity effects, low miRNAs content, etc. question

Inactive Publication Date: 2013-03-20
EAST CHINA NORMAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its detection still has the following defects: the detection sample needs to be processed through cumbersome steps, including enrichment extraction and purification of total RNA in cells, tissues or blood samples, and then reverse transcribed into cDNA, and PCR is required in some cases Amplified before loading and detection
The root cause is that the content of miRNAs in cells, tissues or blood samples is very low, often at the pM-fM level, exceeding the limit low value of current detection methods
Moreover, other complex components in biological samples, such as sugars, lipids, metal ions, macromolecular proteins, etc., will also interfere with the binding of target miRNAs to probes, thus having a significant impact on the sensitivity and specificity of detection
The cumbersome pre-processing process is currently one of the bottlenecks restricting the large-scale clinical use of biochips

Method used

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  • Silicon nanowire chip simultaneously detecting miRNAs and protein markers and detection method and application of silicon nanowire chip
  • Silicon nanowire chip simultaneously detecting miRNAs and protein markers and detection method and application of silicon nanowire chip
  • Silicon nanowire chip simultaneously detecting miRNAs and protein markers and detection method and application of silicon nanowire chip

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0063] Example 1 Preparation of silicon nanowire chips for simultaneous detection of miRNAs and protein markers

[0064] The preparation method and process of the silicon nanowire chip used for simultaneous detection of miRNAs and protein markers in the present invention, such as Figure 4 As shown, as follows:

[0065] Step S01: providing a semiconductor substrate 1;

[0066] Step S02: using a wet oxygen oxidation process to grow a silicon dioxide isolation layer 2 on the semiconductor substrate;

[0067] Step S03: growing a polysilicon layer 3 on the silicon dioxide isolation layer;

[0068] Step S04: using a dry plasma etching process to pattern the polysilicon layer 3 to form a silicon nanowire array 4;

[0069] Step S05: using a thermal oxidation method, grow a passivation layer with a certain thickness (for example: 3nm-15nm) on the silicon nanowire array 4, and grow and form TaN / Ta 2 o 5 layer, wherein the passivation layer structure from top to bottom includes SiO...

Embodiment 2

[0072] Example 2 Simultaneous detection of acute myocardial infarction miRNAs and protein markers

[0073] Using the silicon nanowire chip that can simultaneously detect miRNAs and protein markers of acute myocardial infarction, by modifying different probes or antibodies on the silicon nanowires, the simultaneous detection of miRNAs and protein markers related to acute myocardial infarction can be directly and simultaneously detected in plasma samples Research purposes.

[0074] Its detection method is as follows:

[0075] 1) Each silicon nanowire detects the current value under the voltage of 0-5V (gradient 0.1V) before loading the sample;

[0076] 2) Each silicon nanowire detects the current value under the voltage of 0-5V (gradient 0.1V) after loading the sample;

[0077] 3) Calculate the resistance value of each silicon nanowire before and after sample loading according to the voltage-to-current ratio;

[0078] 4) The resistance value ratio of each silicon nanowire bef...

Embodiment 3

[0082] Embodiment 3 Detection application of the silicon nanowire chip of the present invention

[0083] Because miRNAs and protein markers appear in different phases of changes in plasma samples of patients with myocardial infarction, the present invention prolongs the time window for diagnosis of myocardial infarction, for example, 1-72 hours, especially increases the early (within 2 hours) diagnosis possibility. A 200uL plasma sample was taken and combined with single-stranded DNA probes or antibodies modified on the surface of different silicon nanowires, resulting in changes in the resistance of each silicon nanowire before and after sample loading. Since each silicon nanowire is modified with probes for different miRNAs or antibodies for different protein markers, the absolute value of the concentration of each specific miRNA or protein marker can be obtained by analyzing the change of resistance and comparing with the standard curve. Proceed as follows:

[0084] 1) O...

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Abstract

The invention discloses a silicon nanowire chip simultaneously detecting miRNAs and protein markers. The silicon nanowire chip comprises at least one of integrated configuration of single-chain deoxyribonucleic acid (DNA) probe in a silicon nanowire array, and integrated configuration of protein marker antibody in the silicon nanowire array. The invention further discloses a structure and a preparation method of the silicon nanowire chip, a detection method used for simultaneously detecting the miRNAs and the protein markers, and application in detection of the acute myocardial infarction miRNAs. The silicon nanowire chip simultaneously detecting miRNAs and protein markers and the detection method and the application of the silicon nanowire chip achieve rapid and simultaneous detection of the acute myocardial infarction miRNA and the protein markers, and have the advantages of being high in sensitivity, high in detection speed, easy to integrate, high in flux, high in stability, resistant to pollution, and the like.

Description

technical field [0001] The invention relates to the field of biomolecular detection, in particular to a silicon nanowire chip for simultaneously detecting miRNAs and protein markers, a preparation method, a detection method and an application thereof. Background technique [0002] At present, cardiovascular disease is a common disease that threatens human health with high incidence, while acute myocardial infarction causes a great burden to the society and patients' families because of its rapid onset, high mortality rate and disability rate. If the harm of acute myocardial infarction is to be reduced, early diagnosis, targeted treatment, and adjustment of medication according to the progress of the disease are essential links, and detection and monitoring of certain biomarkers are obviously helpful for the implementation of these aspects. In addition to several protein markers cTnT, cTnI, CK-MM and CK-MB commonly used in clinical practice, microRNAs (miRNAs) are considered ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/04B81C1/00
CPCH01L29/16B82Y10/00G01N33/551B82Y15/00B81C1/00H01L21/02B82Y40/00H01L29/0673
Inventor 赵宇岚朱建军何靖蒋宾
Owner EAST CHINA NORMAL UNIV