Detection method for nucleic acid amplification product

Abstract

The invention provides a detection method for a nucleic acid amplification product, particularly a detection and pollution control method for the nucleic acid amplification reaction product with no need for opening a reaction vessel. The detection method includes steps that: before a nucleic acid amplification reaction, solid-state microcrystalline waxes wrapped with nucleic acid dye are added into the reaction vessel, and the microcrystalline waxes are guaranteed not to contact with reaction liquid; during the nucleic acid amplification reaction, the amplification reaction temperature is maintained to be lower than the melting point of the microcrystalline waxes, and the microcrystalline wax beads are guaranteed not to contact with the reaction liquid; after the nucleic acid amplification reaction, the reaction vessel is heated to the melting point of the microcrystalline waxes which are converted into a liquid state from a solid state and float on the upper layer of the nucleic acid reaction liquid; the wrapped dye enters the reaction liquid, is evenly mixed with the nucleic acid amplification reaction liquid, the nucleic acid amplification product is combined with the dye, and a color reaction is performed.

Description

technical field [0001] The invention relates to an experimental detection method, in particular to a detection method of nucleic acid amplification reaction and its products. Background technique [0002] Biological genetic material nucleic acid can be used as a target molecule for identifying organisms and diagnosing pathogens, but the molecular weight of the target nucleic acid in the sample is usually small, so it is necessary to amplify the target nucleic acid molecule in the sample, and the in vitro synthesis reaction of nucleic acid, related PCR technology, SDA technology and LAMP technology have been invented and applied successively. These technologies have high amplification efficiency, and can efficiently amplify several to dozens of target nucleic acid molecules into a huge amount of nucleic acid molecule products, which can be used for the diagnosis and treatment of various pathogenic microorganisms. Nucleic acid molecular identification of biological samples pro...

AI Technical Summary

Problems solved by technology

[0003] There are many methods for detecting nucleic acid amplification products, some require expensive instruments, and some detection reagents have an inhibitory effect on the amplification reaction, which limits the popularization and application of these methods, such as the LAMP reaction. In view of its high efficiency of nucleic acid synthesis, In the implementation process, the amplification reaction needs to be carried out in a closed reaction container, and the detection of the product cannot adopt the method of opening the container to sample the electrophoresis to identify the amplified product or adding a dye; because if the reaction container is opened, the target nucleic acid is easily produced. The aerosol contamination of product molecules leads to irreversible and continuous false positive results in subsequent detection experiments. Once such product contamination of the experimental environment occurs, the contamination is extremely difficult to eliminate in a short period of time, resulting in the same LAMP targeting a certain target. The detection experiment cannot be carried out for a long time, which is the biggest problem in the application of LAMP to detect pathogens, which greatly limits the popularization and application of LAMP technology

[0004] Although there is also a method of judging the detection of nucleic acid amplification reaction by observing the generation of turbidity without opening the container, under the visual conditions of different experimenters, the judgment of whether the turbidity is generated or not is often inconsistent, and expensive instruments must be used to ensure Its precision is difficult to popularize at the grassroots level

And if the dye is added before the reaction, it will interfere with the nucleic acid synthesis reaction

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