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SPR (surface plasma resonance) Sensor for detecting HIV (human immunodeficiency virus) related genes and preparation and application thereof

A sensor and gene technology, applied in the field of ion resonance sensing, can solve the problems of poor HIV detection specificity, long detection time, cumbersome process, etc.

Inactive Publication Date: 2017-10-17
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the shortcomings of the prior art described above, the purpose of the present invention is to provide a SPR sensor for detecting HIV-related genes and its preparation and application, which is used to solve the problem of poor detection specificity, cumbersome process, and time-consuming detection of HIV in the prior art. long wait question

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  • SPR (surface plasma resonance) Sensor for detecting HIV (human immunodeficiency virus) related genes and preparation and application thereof
  • SPR (surface plasma resonance) Sensor for detecting HIV (human immunodeficiency virus) related genes and preparation and application thereof
  • SPR (surface plasma resonance) Sensor for detecting HIV (human immunodeficiency virus) related genes and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Example 1 Preparation of surface plasmon resonance HIV-related gene detection sensor

[0112] 1. Materials and methods

[0113] 1.1 Material

[0114] The DNA sequence purified by HPLC was synthesized by Shanghai Shenggong Biological Engineering Co., Ltd. TE buffer (10mM TrisHCl, 1mM EDTA, pH 8.0) was purchased from Shanghai Shenggong Bioengineering Co., Ltd., DNA marker was purchased from Dalian Takara Company, KH 2 PO 4 , NaCl, MgCl 2 , Na 3 PO 4 ·12H 2 O, EDTA-2Na and other reagents were purchased from Chongqing Maoye Chemical Reagent Co., Ltd.

[0115] 1.2 Testing equipment

[0116] The Biocore X surface plasmon resonance instrument is a product of Biocore AB, Sweden.

[0117] 1.3 Detection principle

[0118] Such as Picture 10 As shown, in the homogeneous reaction system, the target sequence T triggers the entropy-driven chain displacement cyclic amplification reaction through the foothold of the Q chain, freeing the P single chain in the three-strand complex PRQ to form the ...

Embodiment 2

[0136] Example 2 Verification of the feasibility of a surface plasmon resonance HIV-related gene detection sensor

[0137] 1. The characterization of the entropy-driven strand displacement cycle amplification system, double-layer DNA tetrahedral nano-molecules and regeneration solution NaOH successively.

[0138] The injection volume of the capture probe is 100 μL, and the molar concentration is 1 μM.

[0139] The number of Q / C base complementary pairings is 6, and the Q chain sequence is Q6:

[0140] 5'-ATGTGGAAAATCTCTAGCAGTAGGGCCGTAAGTTAGTTGGAGACGTAGGCGGATCC-3' (SEQ ID NO. 4). After hybridization solution, triple-strand complex PRQ, single-strand F, and target sequence T are mixed to obtain a mixed solution, the concentration of triple-strand complex PRQ and single-strand F in the mixture is 250 nM, and the concentration of target sequence T is 100 nM, which is driven by entropy The chain displacement cycle amplification reaction temperature is 37°C, the reaction time is 30min, and...

Embodiment 3

[0171] Example 3 Surface plasmon resonance HIV-related gene detection sensor and its use condition influence experiment

[0172] We also determined several important conditions in the present invention (namely, the number of Q / C base complementary pairings, the entropy-driven strand displacement cycle amplification reaction time, the reaction temperature, and the four measurement conditions of the double-layer DNA tetrahedral nanomolecule injection volume. ) Further experiments were carried out on the influence of the test results. For each condition, five points were selected from low concentration to high concentration to conduct a series of experiments.

[0173] 1. In order to investigate the influence of the number of Q / C base pairings on the surface plasmon resonance HIV-related gene detection sensor, this experiment used different base pairing numbers to construct the surface plasmon resonance sensor. Such as Figure 4 It can be seen that the signal-to-noise ratio varies wit...

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Abstract

The invention provides an SPR (surface plasma resonance) sensor for detecting HIV (human immunodeficiency virus) related genes and preparation and application thereof; the preparation of the SPR sensor comprises: preparing an entropy-driven strand displacement amplifying system, including, (a), preparing a probe, to be specific, using DNA single strands P, R and Q to prepare three-strand composite PRQ, (b), taking the three-strand composite PRQ obtained in step (a), mixing hybridization solution, the three-strand composite PRQ, single strand F and target sequence T, and hatching to obtain a reaction liquid, (c), fixing a capture probe to the surface of a working chip, inserting the working chip in a surface plasma resonance apparatus, and injecting the reaction liquid of step (b) as a sample to the surface of the working chip wherein the capture probe is fixed so as to obtain the entropy-driven strand displacement amplifying system. The surface plasma resonance sensor and detection system suitable for detecting HIV related genes are successfully constructed herein; by using the sensor of the invention to measure HIV related gears, good sensitivity, stability and reproducibility are achieved.

Description

Technical field [0001] The invention relates to the field of nucleic acid detection and surface plasmon resonance sensing, in particular to an SPR (surface plasmon resonance) sensor for detecting HIV (human immunodeficiency virus) related genes and its preparation and application. Background technique [0002] Since the first case of HIV infection was discovered in mainland China in 1985, the global epidemic of AIDS has become increasingly serious and the situation is grim. According to UNAIDS data, 1.1 million people died of AIDS globally in 2015, and 2.1 million new HIV infections were added that year. The AIDS epidemic has brought great damage to human health and social economy. The main purpose of human immunodeficiency virus (HIV) testing is to diagnose HIV pathogens, guide clinical medications, and detect HIV carriers. Early detection and intervention can help control the disease and block the route of transmission in time. At present, most of the diagnosis of HIV infecti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/682C12Q1/703C12Q2565/628C12Q2531/119C12Q2565/519
Inventor 颜玉蓉丁世家程伟杨建儒刁玮程文彬晏小玉马洪敏
Owner CHONGQING MEDICAL UNIVERSITY
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