Mercuric ion detection kit based on constant-temperature cascading nucleic acid amplification and detection method thereof

Abstract

The invention provides a mercuric ion detection kit based on constant-temperature cascading nucleic acid amplification and a detection method thereof. Thymine-containing DNA identification elements are specifically bound with mercuric ions and then are folded to form an intra-molecular stem-loop structure; The 3' terminal of the DNA can be identified by polymerase, the DNA itself serves as a template to start strand displacement amplification reaction so as to produce a large amount of single-stranded DNA products; the single-stranded DNA products can open a restriction enzyme cutting site containing molecular beacon stem-loop structure to produce fluorescence signals; at the same time, the single-stranded DNA products can be also used as primers, and hybridization combined molecular beacons are used as templates to trigger secondary strand displacement amplification reaction, and released SDA products can form heteroduplexes with new molecular beacons so as to produce cascaded-amplified fluorescent signals. The detection method is high in sensitivity, ingeniously achieves cascading amplification of strand displacement amplification reaction without system complexity increase, is high in amplification efficiency and response speed and can achieve mercuric ion quantification within 30 minutes, and the detection limit is as low as 2 nM.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to the quantitative detection of mercury ions, in particular to a mercury ion detection kit based on constant temperature cascade nucleic acid amplification and a detection method thereof. technical background [0002] Heavy metal ions have high stability in the environment and are difficult to be degraded by microorganisms. Once they enter the environment, they are difficult to be repaired naturally, and can be continuously enriched and passed on in the ecosystem. Even a small amount of ingestion can cause great toxicity, which is a major threat to ecological balance and human health. As a typical representative of heavy metal ion pollution, the accumulation of mercury ions in biological tissues causes DNA damage, affects ligand-receptor interactions, destroys the immune system, and causes a series of diseases, such as brain damage, kidney failure, various cognition and movement disorder...

AI Technical Summary

Problems solved by technology

[0006] In order to overcome the defects in the above-mentioned prior art, the object of the present invention is to provide a mercury ion detection kit and its detection method based on constant temperature cascade nucleic acid amplification, so as to overcome the insufficient sensitivity, complicated design and long time-consuming of the prior art. shortcoming

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