Method for detecting saxitoxin on basis of embedded dye and DNA interaction

A technology for saxitoxin and a detection method, which is applied in the field of detection and research of saxitoxin, can solve the problems of high detection cost, complicated optical signal sequence and ineffective effect, and difficult to obtain instruments, and achieves simple detection method, overcoming Effects of Immobilization and Detection Platform Construction Complexity

Inactive Publication Date: 2017-09-19
HEFEI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The reason is mainly that the length of the two sequences is relatively long, and the secondary structure is relatively complex, which makes it difficult to carry out replacement, amplification, and fixation using base pairing and elimination, and the optical signal generated by using conformational changes is due to the sequence itself. The effect of complexity is not obvious
[0006] At present, there are only BLI and electrochemical analysis methods for the construction of DNA-targeted saxitoxin sensors. These methods use surface-interface analysis instruments to immobilize DNA probes and then monitor the detection process in real time. They are strongly dependent on instruments and have a long cycle. The detection cost is high, and the equipment is difficult to obtain, so it is necessary to find a universal method to design the developed sequence to achieve the purpose of application

Method used

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  • Method for detecting saxitoxin on basis of embedded dye and DNA interaction
  • Method for detecting saxitoxin on basis of embedded dye and DNA interaction

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Effect test

Embodiment 1

[0023] Firstly, 1 μM DNA was dissolved in 10 mM Tris-HCl buffer solution with pH=7 and quenched, then saxitoxin was added to the Tris-HCl buffer solution to interact with DNA, and the interaction time was 25 min. After the time for the interaction is up, add 1 μM fluorescent violet to the Tris-HCl buffer, wait for 25 minutes, use a fluorescence spectrophotometer to measure the fluorescence spectrum, and calculate the stone by comparing the peak at 652 nm of the measured spectrum with the standard curve. Toxin concentration of saxitoxin.

Embodiment 2

[0025] First, 1 μM DNA was dissolved in 10 mM Tris-HCl buffer at pH=7 and quenched, then saxitoxin was added to the Tris-HCl buffer to interact with DNA, and the interaction time was 35 min. After the time for the interaction is up, add 1 μM fluorescent violet to the Tris-HCl buffer, wait for 35 minutes, use a fluorescence spectrophotometer to measure the fluorescence spectrum, and calculate the stone by comparing the peak at 652 nm of the measured spectrum with the standard curve. Toxin concentration of saxitoxin.

Embodiment 3

[0027] Firstly, 1 μM DNA was dissolved in 10 mM Tris-HCl buffer solution with pH=7 and quenched, and then saxitoxin was added to the Tris-HCl buffer solution to interact with DNA, and the interaction time was 30 min. After the time for the interaction is up, add 1 μM fluorescent violet to the Tris-HCl buffer, wait for 30 minutes, use a fluorescence spectrophotometer to measure the fluorescence spectrum, and calculate the stone by comparing the peak at 652 nm of the measured spectrum with the standard curve. Toxin concentration of saxitoxin.

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Abstract

The invention discloses a method for detecting saxitoxin on the basis of embedded dye and DNA interaction. The method comprises the following specific steps: dissolving DNA in a buffer solution at first, then adding the saxitoxin in the buffer solution, carrying out interaction on the saxitoxin and DNA for 25-35 minutes, when interaction time is up, adding the embedded dye in the buffer solution, waiting for 25-35 minutes, then carrying out fluorescence spectrometry, and calculating the concentration of the saxitoxin through the peak value of the measured spectrum according to a standard curve. The concentration of the saxitoxin can be measured quantitatively through measurement to fluorescent light, complicated instruments and construction of a detecting platform are not required, the detection mode is simple, rapid and accurate, therefore, the technological difficulty that traditional measurement depends on complicated and high-cost instruments is avoided, and complexity on fixing of DNA and construction of the detecting platform is avoided.

Description

technical field [0001] The invention relates to the detection and research of saxitoxin, in particular to a detection method of saxitoxin based on the interaction between embedded dye and DNA. Background technique [0002] Paralytic shellfish toxin is a kind of neurotoxin, which is extremely toxic. It can hinder the conduction of nerve impulses and inhibit the central nervous system, resulting in paralysis of the respiratory system and death. Among them, the most toxic saxitoxin can cause death within 15 minutes. Since it cannot be destroyed by digestive enzymes and is colorless and odorless, its detection and research are particularly important. The traditional mouse detection method and post-HPLC fluorescence analysis detection technology cannot be widely used in daily life due to limitations in animal experiments and large-scale instruments. All kits currently on the market are antibody-based protein kits. Due to the limitations of its stability and large-scale preparat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6486
Inventor 郑斌程盛朱金苗朱文娟朱艺
Owner HEFEI NORMAL UNIV
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