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Detection technology combining multi-cross displacement amplification with gold nano detection and application of detection technology

A technology of cross-replacement and detection technology, which can be used in microorganism-based methods, microorganism determination/inspection, biochemical equipment and methods, etc. It can solve specific problems and achieve the effect of convenient methods.

Pending Publication Date: 2020-02-21
首钢医院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the widely used constant temperature amplification techniques include rolling circle amplification (RCA), strand displacement amplification (SDA), helicase-dependent constant temperature amplification (HDA), loop-mediated constant temperature amplification (LAMP) and crossover. Amplification (CPA), etc., there are not enough convenient, fast, sensitive and specific problems in the detection

Method used

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  • Detection technology combining multi-cross displacement amplification with gold nano detection and application of detection technology
  • Detection technology combining multi-cross displacement amplification with gold nano detection and application of detection technology
  • Detection technology combining multi-cross displacement amplification with gold nano detection and application of detection technology

Examples

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Effect test

Embodiment 1

[0105] This embodiment provides a detection technology combining multiple cross-displacement amplification and gold nanometer detection, including:

[0106] From the 3' end of the target gene fragment, set the first arbitrary sequence F1s, the second arbitrary sequence P1s, from the 5' end of the target gene fragment, set the third arbitrary sequence F2, and the fourth arbitrary sequence P2, setting a fifth arbitrary sequence C1 at the 5' end of the second arbitrary sequence P1s, and / or setting a sixth arbitrary sequence C2s at the 3' end of the fourth arbitrary sequence P2;

[0107] Provide a replacement primer F1, the primer F1 contains a sequence complementary to the sequence F1s, provide a cross primer CP1, the primer CP1 sequentially contains a sequence C1s complementary to the sequence C1 and a sequence P1 from the 5' end, and provide a replacement primer F2, the The primer contains a sequence complementary to the sequence F2s, and a cross primer CP2 is provided, and the...

Embodiment 2

[0117] This embodiment provides a detection technology combining multiple cross-displacement amplification and gold nanometer detection, including:

[0118] From the 3' end of the target gene fragment, set the first arbitrary sequence F1s, the second arbitrary sequence P1s, from the 5' end of the target gene fragment, set the third arbitrary sequence F2, and the fourth arbitrary sequence P2, setting a fifth arbitrary sequence C1 at the 5' end of the second arbitrary sequence P1s, and / or setting a sixth arbitrary sequence C2s at the 3' end of the fourth arbitrary sequence P2;

[0119] Provide a replacement primer F1, the primer F1 contains a sequence complementary to the sequence F1s, provide a cross primer CP1, the primer CP1 sequentially contains a sequence C1s complementary to the sequence C1 and a sequence P1 from the 5' end, and provide a replacement primer F2, the The primer contains a sequence complementary to the sequence F2s, and a cross primer CP2 is provided, and the...

Embodiment 3

[0129] This embodiment provides a detection technology combining multiple cross-displacement amplification and gold nanometer detection, including:

[0130] From the 3' end of the target gene fragment, set the first arbitrary sequence F1s, the second arbitrary sequence P1s, from the 5' end of the target gene fragment, set the third arbitrary sequence F2, and the fourth arbitrary sequence P2, setting a fifth arbitrary sequence C1 at the 5' end of the second arbitrary sequence P1s, and / or setting a sixth arbitrary sequence C2s at the 3' end of the fourth arbitrary sequence P2;

[0131] Provide a replacement primer F1, the primer F1 contains a sequence complementary to the sequence F1s, provide a cross primer CP1, the primer CP1 sequentially contains a sequence C1s complementary to the sequence C1 and a sequence P1 from the 5' end, and provide a replacement primer F2, the The primer contains a sequence complementary to the sequence F2s, and a cross primer CP2 is provided, and the...

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Abstract

The invention belongs to the technical field of biology and particularly relates to a detection technology combining multi-cross displacement amplification with gold nano detection and application ofthe detection technology in gene detection. According to the detection technology combining multi-cross displacement amplification with gold nano detection, by amplifying a label labelled at the 5' end of a primer C1 in multi-cross displacement amplification, the method is directed to the feature that amplification products of a specific gene pgaD (labeling isothiocyanate fluorescein FITC) of acinetobacter baumannii and a carbapenem drug resistance gene blaOXA-23-like (labeling digoxigenin Dig) can be visually detected by a gold nano-biosensor; and the method is convenient, fast, sensitive andspecific and is suitable for being popularized and applied to detection of various specific nucleotide fragments and being applied to detection of pathogenic bacteria corresponding to the specific nucleotide fragments.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a detection technology combined with multi-cross displacement amplification and gold nanometer detection and its application. Background technique [0002] Nucleic acid detection, that is, nucleic acid amplification detection technology generally refers to the detection technology of screening specific genes by means of amplifying DNA or RNA. Nucleic acid amplification technology includes PCR amplification technology, constant temperature amplification technology, etc. Compared with PCR and its derivative technologies, it does not rely on thermal cycle amplification equipment, the entire amplification process is constant at a fixed temperature, has the characteristics of fast reaction speed, strong sensitivity, and high specificity, which is conducive to rapid amplification and convenient detection and on-site diagnosis. At present, the widely used constant temperature a...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/6804C12Q1/04C12R1/01
CPCC12Q1/689C12Q1/6844C12Q1/6804
Inventor 胡守奎闫琳琳赵帆牛莉娜蔡煜吴蕾朱晓雪高乃姝农金轻邢喆
Owner 首钢医院有限公司
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