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LSPR (localized surface plasma resonance) sensing device and preparation method thereof as well as DNA detection method

A technology of a sensing device and a manufacturing method, applied in the field of biochemical detection, can solve the problems of single incident light and lack of sensitivity, achieve the effects of accurate detection, avoid wave peak broadening, and improve accuracy and sensitivity

Inactive Publication Date: 2014-10-01
镇江威新生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional LSPR sensor uses a relatively consistent symmetrical structure, which produces a single absorption peak for incident light. Although these symmetrical structures have satisfied the detection of most biological macromolecules, for small biological molecules such as bacterial DNA still lacks sufficient sensitivity

Method used

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  • LSPR (localized surface plasma resonance) sensing device and preparation method thereof as well as DNA detection method
  • LSPR (localized surface plasma resonance) sensing device and preparation method thereof as well as DNA detection method
  • LSPR (localized surface plasma resonance) sensing device and preparation method thereof as well as DNA detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] In the present invention, select Gram-positive bacteria to carry out the present invention's operation as bacterial standard sample:

[0090] S11, take 0.5-2ml of the cultured bacteria solution in the EP tube, centrifuge at high speed for 30s, and discard the supernatant; add buffer RB to the EP tube to resuspend, and discard the supernatant after high-speed centrifugation; repeat this step 2 to 3 times;

[0091] S12, add an appropriate amount of lysozyme, mix upside down, and incubate at 37°C for 30-60 minutes; after high-speed centrifugation, discard the supernatant and resuspend the cells in buffer RB by shaking or pipetting;

[0092] S13, add 20 μl of RNase A (25 mg / ml) solution to the solution obtained in step S13, shake and mix, and place at room temperature for 5-10 minutes;

[0093] S14, add the binding solution CB, and then add a small amount of isopropanol, at this time there will be flocculent sediment, immediately vortex and mix thoroughly to obtain a mixtur...

Embodiment 2

[0122] Based on the above comparison, in Example 2, the DNA extracted from Staphylococcus aureus is used as the DNA to be tested to carry out the steps in Example 1, and the DNA capture probe still uses the 16srRNA of Gram-positive bacteria in Example 1 The capture probe sequence of the gene; the detection step is carried out according to the same steps as in Example 1.

[0123] In the final absorption spectrum, its absorption peak obtained does not move, then it is because the probes fixed on the surface of the DNA to be measured and the LSPR sensor (that is, the quartz glass plate of the above-mentioned asymmetrical XI nano-metal structure) cannot be combined, That is to say, the detected bacteria are not the bacteria corresponding to the probe.

[0124] Of course, in the repeated implementation of the above embodiment, there may be a result that the absorption peak moves to the left, because it indicates that the fixed probe falls off, causing the absorption peak to move to t...

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Abstract

The invention provides an LSPR (localized surface plasma resonance) sensing device. The LSPR sensing device comprises a substrate, wherein the substrate is provided with a metal layer, the metal layer is in an XI shape, and the size of XI is of a nano grade. The LSPR sensing device prepared through the method establishes an asymmetric XI nano gold structure, a specific peak in the LSPR phenomenon can be produced independently by the X and I structure; after the XI structure is formed by combining the X structure and I structure, the spectrum of the two structures can be collectively heterozygozed, and a multipeak phenomenon occurs; the XI structure can have a coupling effect, so that the heterozygosis peak is deviated; the DNA detection precision and sensitivity can be greatly improved; the noise caused by the nonuniform structure caused by a traditional method can be avoided, the situation that the peak is widened due to the disorder structure can be avoided, the heterozygosis of the specific spectrums of the two structures is resonated with Fano produced by the asymmetric structure, so that the LSPR sensor is more accurate in detection.

Description

technical field [0001] The invention belongs to the technical field of biochemical detection, and in particular relates to an LSPR sensing device based on localized surface plasmon resonance technology, a preparation method thereof, and a DNA detection method. Background technique [0002] At present, it is usually more accurate and feasible to use molecular biology methods to detect bacteria. For example, the commonly used PCR method and gene chip are all through extracting the DNA of the bacteria to be tested, and then testing and verifying the DNA, so that the results of the bacteria to be tested can be known. [0003] Among them, the PCR method and gene chip that are usually used; the PCR method uses the DNA extracted from the bacteria to amplify the bacterial genes in large quantities, and in the PCR process, the amplified product is detected by a gene probe specifically designed based on the target gene of the bacteria. In order to achieve bacterial detection; the imp...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12Q1/68
CPCC12Q1/6825G01N21/554C12Q2565/628
Inventor 李科铮吴宇亮于洪宇
Owner 镇江威新生物科技有限公司
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