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A detection method for nucleic acid amplification

A detection method, nucleic acid technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems affecting the accuracy of detection results, etc.

Inactive Publication Date: 2020-08-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, relying on these devices, the uniformity of the sample during the dilution process still has a great impact on the test results. If the sample is not mixed evenly, it will directly affect the accuracy of the test results.

Method used

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  • A detection method for nucleic acid amplification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: The method of using the visualized nucleic acid amplification detection device—using antigen-modified primers for test strip detection

[0018] 1) in such as figure 1 Add nucleic acid amplification reaction solution and antigen modification primers to PCR tube 1 shown, and add MnO to PCR tube 2 2 and the mixed solution of calcein; PCR tube 3 is added with pyrophosphate (sodium pyrophosphate or potassium pyrophosphate) solution.

[0019] 2) Connect tube 1, tube 2, and tube 3 of the PCR tube to the three connection ports of the visualized nucleic acid amplification detection device 4 respectively, and seal them. Nucleic acid detection test strips 6 are pre-installed in the detection chamber 5 of the visualized nucleic acid amplification detection device.

[0020] 3) Place the sealed device on a PCR instrument or a metal bath for amplification. The three PCR tubes filled with reagents are just placed in the metal bath or the heating hole of the PCR instrument...

Embodiment 2

[0025] Example 2: The method of use of the device - using antigen-modified probes for test strip detection 1

[0026] 1) in such as figure 1 The nucleic acid amplification reaction solution is added to the PCR tube 1 shown, and MnO is added to the PCR tube 2 2 and the mixed solution of calcein and antigen-modified probe; add pyrophosphate (sodium pyrophosphate or potassium pyrophosphate) solution to PCR tube 3.

[0027] Remaining steps are with embodiment 1.

Embodiment 3

[0028] Example 3: The method of using the device—using antigen-modified probes for test strip detection 2

[0029] 1) in such as figure 1 The nucleic acid amplification reaction solution is added to the PCR tube 1 shown, and MnO is added to the PCR tube 2 2 mixed solution with calcein and antigen-modified probe 1; PCR tube 3 is added with pyrophosphate (sodium pyrophosphate or potassium pyrophosphate) solution and antigen-modified probe 2.

[0030] Remaining steps are with embodiment 1.

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Abstract

The invention provides a detection method for nucleic acid amplification. The detection method comprises the following steps: adding a reaction amplification solution in a first PCR pipe, then adding calcein and a MnO2 nano-material into a second PCR pipe and adding a pyrophosphate solution into a third PCR pipe; respectively placing test strip probes into the second PCR pipe and the third PCR pipe, or adding an antigen-modified primer into the first PCR pipe; with a visible nucleic acid amplification detection apparatus with three interfaces as a cover, respectively and hermetically connecting the PCR pipes with the three interfaces of the visible nucleic acid amplification detection apparatus, and after completion of reaction, mixing the reaction amplification solution, calcein, MnO2 and the pyrophosphate solution. According to the invention, uniform mixing can be determined by determining whether the color of mixed liquor is yellow green, and then test strips are used for detecting the constant-temperature amplification results of nucleic acid; so the degree of uniform mixing of a sample before detection can be monitored without opening of the cover, and more accurate results are obtained through detection with the test strips.

Description

technical field [0001] The invention belongs to the field of nucleic acid analysis and detection, and in particular relates to a visual detection method for nucleic acid amplification products. Background technique [0002] Nucleic acid amplification technology is of great significance in the field of analysis and detection today. Food safety testing, environmental monitoring, medical diagnosis, forensic identification and other fields all rely on nucleic acid amplification technology. The earliest and most classic nucleic acid amplification technology is to achieve template denaturation, primer annealing and extension under the action of heat-resistant polymerase, relying on continuous cycling of temperature to achieve a large amount of accumulation of target products. This technology is called polymerization. Enzyme chain reaction (PCR). However, the biggest defect of PCR is that its constant temperature change process puts forward relatively high requirements on the ins...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2527/125C12Q2563/107C12Q2565/625
Inventor 吴坚王柳应义斌
Owner ZHEJIANG UNIV