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Method for detecting poly ADP-ribose polymerase-1 with gold nanorod probe

A technology of polyadenosine diphosphate-ribose and gold nanorods, which is applied in the field of biosensing, can solve the problems of complicated operation procedures, narrow detection range and high cost, and achieve the effect of low operating cost, low raw material cost and simple principle

Active Publication Date: 2020-06-02
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The radioisotope labeling method requires prior labeling of the substrate NAD + , the detection result is more reliable and sensitive, and can detect a very small amount of PARP-1, but the detection range is narrow, the operation procedure is complicated, the cost is high, it requires very precise measuring instruments, it takes a long time, and there are problems in the detection of a large number of actual samples. more restrictive conditions

Method used

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  • Method for detecting poly ADP-ribose polymerase-1 with gold nanorod probe
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  • Method for detecting poly ADP-ribose polymerase-1 with gold nanorod probe

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Embodiment 1

[0027] The analytical method for detecting poly ADP-ribose polymerase-1 by gold nanorod probe colorimetric method based on electrostatic interaction, the detection steps are:

[0028] Synthesis steps of gold nanorod probes: gold nanorods were synthesized by a one-pot method. Take a mouth bottle, add 38mL of CTAB solution with a concentration of 0.2mol / L to 30mL of deionized water, then quickly add 3mL of chloroauric acid dropwise, and then slowly add 346μL of 0.02mol / L silver nitrate solution and 2mL containing 0.044g After the hydroquinone solution was reacted for 5 minutes, 2.6 mL of freshly prepared sodium borohydride ice water solution with a concentration of 0.5 mmol / L was added, shaken evenly, and incubated at 30°C for more than 12 hours, and the total volume was 76 mL. Finally, the probe solution was centrifuged at 13000 rpm at 20° C. for 20 to 30 minutes, redissolved in deionized water after centrifugation twice, and stored at room temperature. This solution was used ...

Embodiment 2

[0033] The analytical method for detecting poly ADP-ribose polymerase-1 by gold nanorod probe colorimetric method based on electrostatic interaction, the detection steps are:

[0034] Synthesis steps of gold nanorod probes: gold nanorods were synthesized by a one-pot method. Take a mouth bottle, add 38mL of CTAB solution with a concentration of 0.2mol / L to 30mL of deionized water, then quickly add 3mL of chloroauric acid dropwise, and then slowly add 346μL of 0.02mol / L silver nitrate solution and 2mL containing 0.044g After the hydroquinone solution was reacted for 5 minutes, 2.6 mL of freshly prepared sodium borohydride ice water solution with a concentration of 0.5 mmol / L was added, shaken evenly, and incubated at 30°C for more than 12 hours, and the total volume was 76 mL. Finally, the probe solution was centrifuged at 13000 rpm at 20° C. for 20 to 30 minutes, redissolved in deionized water after centrifugation twice, and stored at room temperature. This solution was used ...

Embodiment 3

[0039] The analytical method for detecting poly ADP-ribose polymerase-1 by gold nanorod probe colorimetric method based on electrostatic interaction, the detection steps are:

[0040] Synthesis steps of gold nanorod probes: gold nanorods were synthesized by a one-pot method. Take a mouth bottle, add 38mL of CTAB solution with a concentration of 0.2mol / L to 30mL of deionized water, then quickly add 3mL of chloroauric acid dropwise, and then slowly add 346μL of 0.02mol / L silver nitrate solution and 2mL containing 0.044g After the hydroquinone solution was reacted for 5 minutes, 2.6 mL of freshly prepared sodium borohydride ice water solution with a concentration of 0.5 mmol / L was added, shaken evenly, and incubated at 30°C for more than 12 hours, and the total volume was 76 mL. Finally, the probe solution was centrifuged at 13000 rpm at 20° C. for 20 to 30 minutes, redissolved in deionized water after centrifugation twice, and stored at room temperature. This solution was used ...

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Abstract

The invention discloses a method for detecting poly(ADP-ribose) polymerase-1 (PARP-1) by a gold nano-rod probe colorimetry. The method comprises the following steps: (1) carrying out selection and hybridization of activated double-chain DNA; (2) carrying out synthesis of a gold nano-rod probe; (3) carrying out mixed reaction on the activated double-chain DNA, PARP-1 and NAD+, and preparing poly(ADP-ribose) polymerase polymer (PAR); (4) detecting a reaction solution under the effect of the gold nano-rod probe and the poly(ADP-ribose) polymerase polymer (PAR) and by an ultraviolet-visible spectrograph; and (5) carrying out characterization on a gold rod solution before and after agglomeration by a transmission electron microscope and dark-field light scattering. After self-assembling of theelectropositive gold nano-rod on the surface of electronegative PAR, the solution becomes almost colorless from deep reddish brown, color change is obvious, and therefore, PARP-1 can be detected by the colorimetry. The method has the advantages of low running cost, rapid detection, simplicity and convenience, high sensitivity, good selectivity and the like.

Description

technical field [0001] The invention relates to a technique for quantitatively detecting polyadenosine diphosphate-ribose polymerase-1, belonging to the technical field of biosensing. Background technique [0002] The traditional main methods for detecting PARP-1 include radioactive isotope labeling, western blot, and enzyme-linked immunosorbent assay. The radioisotope labeling method requires prior labeling of the substrate NAD + , the detection result is more reliable and sensitive, and can detect a very small amount of PARP-1, but it has a narrow detection range, complicated operation procedures, high cost, requires very sophisticated measuring instruments, takes a long time, and has problems in the detection of a large number of actual samples. More restrictions. Colorimetry is not only cheap, does not require complex instruments and equipment, but also is simple and fast to measure, accompanied by color changes that can be observed by the naked eye, so it has develope...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/25
Inventor 卫伟吴霜霜刘松琴
Owner SOUTHEAST UNIV
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