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Visual nucleic acid detection method for regulating polymer temperature-sensitive phase change based on nucleic acid

A detection method and polymer technology, which is applied in the field of biomedical detection, can solve the problems of complex detection process, time-consuming, and expensive equipment for nucleic acid detection, and achieve high detection efficiency, low detection cost, and high detection accuracy.

Inactive Publication Date: 2021-10-19
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of the prior art, the purpose of the present invention is to overcome the deficiencies of the prior art, and provide a visual nucleic acid detection method based on nucleic acid-regulated temperature-sensitive phase transition of polymers, which overcomes the dependence on expensive equipment for nucleic acid detection, which needs to be tested in the laboratory It can only be completed with the cooperation of professionals, and the detection process is complicated, time-consuming, and the detection results are not intuitive enough and a series of difficulties

Method used

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  • Visual nucleic acid detection method for regulating polymer temperature-sensitive phase change based on nucleic acid
  • Visual nucleic acid detection method for regulating polymer temperature-sensitive phase change based on nucleic acid
  • Visual nucleic acid detection method for regulating polymer temperature-sensitive phase change based on nucleic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] In this embodiment, the test of sample N:P=32:1

[0029] A mixture of PG1AM (0.1mg / mL) with an amino concentration of 14μM and DNA (18bp) with a phosphate group concentration of 467nM was prepared with ultrapure water, and a sample of N:P=32:1 was obtained after thorough mixing.

[0030] Put the sample N:P=32:1 aqueous solution into a quartz cuvette, and record the turbidity curve at 25-60°C by changing the temperature (0.5°C / min) with a UV-visible spectrophotometer.

Embodiment 2

[0032]This embodiment is basically the same as Embodiment 1, especially in that:

[0033] In this embodiment, the test of sample N:P=2:1

[0034] Prepare a mixture of PG1AM (0.1 mg / mL) with an amino concentration of 14 μM and DNA (18 bp) with a phosphate group concentration of 7 μM with ultrapure water, and mix thoroughly to obtain a sample with N:P=2:1.

[0035] Put the sample N:P=2:1 aqueous solution into a quartz cuvette, and record the turbidity curve at 25-60°C by changing the temperature (0.5°C / min) with a UV-visible spectrophotometer.

Embodiment 3

[0037] This embodiment is basically the same as the previous embodiment, and the special features are:

[0038] In this example, the temperature-sensitive phase transition of the nucleic acid amplification system

[0039] The DNA was first amplified by RCA, and a PG1AM sample with a concentration of 0.1 mg / mL was prepared with ultrapure water. Among them, sample 1 is PG1AM control group, sample 2 is PG1AM and DNA amplified by RCA, and sample 3 is PG1AM and DNA not amplified by RCA.

[0040] Put the sample aqueous solution into a glass bottle, place it in a constant temperature water bath, and take pictures by changing the temperature of the system to record the relevant temperature-sensitive phase transition phenomenon.

[0041] The above-mentioned embodiments are based on a new nucleic acid visual detection method based on nucleic acid-regulated temperature-sensitive phase transition of polymers. After the nucleic acid is combined with a specific polymer, the overall soluti...

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Abstract

The invention discloses a novel nucleic acid visual detection method based on temperature-sensitive phase change of a nucleic acid regulation polymer, and relates to the technical field of biomedical detection, after nucleic acid is combined with a specific polymer, the whole solution shows obvious transmittance change along with temperature change, and the transmittance change is in negative correlation with the nucleic acid concentration in a system. Nucleic acid concentration is taken as a horizontal coordinate, transmittance is taken as a vertical coordinate, and a relation curve of nucleic acid concentration and transmittance, namely a standard curve, is obtained through fitting. During detection, after a certain volume of nucleic acid is mixed with the polymer, the concentration of the nucleic acid in a system can be determined by comparing the transmittance at a specific temperature with the standard curve, so that a novel method for visual detection of the nucleic acid is provided. The method is high in detection efficiency, low in detection cost and high in detection precision.

Description

technical field [0001] The invention relates to the technical field of biomedical detection, in particular to a nucleic acid visual detection method based on nucleic acid regulation of temperature-sensitive phase transition behavior of polymers. Background technique [0002] The rapid detection of nucleic acid is of great significance in modern medical detection, and it is a key means of monitoring and diagnosing some epidemic diseases. Compared with immunological detection, nucleic acid detection has higher accuracy and timeliness: on the one hand, it can be used for pathogen identification and early warning of the intermediate medium of disease transmission, which is indispensable in disease prevention and transmission control; On the one hand, it can make accurate judgments in the early stages of the patient's course of disease, even before there is an obvious immune response, and guide early intervention and precise treatment of the disease, which is the key to saving pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/3151G01N2021/3155
Inventor 张阿方闵泽奇冯乐天徐碧漪李文
Owner SHANGHAI UNIV
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