Molecular detecting method based on exonuclease and G tetramer and detecting kit

Abstract

The invention discloses a molecular detecting method based on an exonuclease and a G tetramer and a detecting kit. Molecules are interacted with a nucleic acid aptamer, chain replacement reaction is started, a stem-loop structural DNA is opened, and then, an exonuclease mediated signal amplification technology is combined, so that a great number of G tetramer structures with activity can be generated. G tetramers are combined with hemin so as to have the catalytic activity of HRP and be capable of catalytically oxidizing TMB to generate blue substrates, a result can be seen by naked eyes, and the concentrations of molecules are directly related to the depth of blue. The molecular detecting method is simple in operation, capable of finishing the whole reaction at the room temperature, relatively high in sensitivity and favorable in specificity and can be used for rapidly detecting molecules on site; the detection limit for 17b-estradiol is 1 pM; and the detected result can be directly seen with naked eyes, but no detecting instruments are used.

Description

technical field [0001] The invention relates to a molecular detection method, in particular to a molecular detection method and a detection kit based on exonuclease and G tetramer. Background technique [0002] There are many harmful molecules in the environment, and it is very practical to detect these harmful small molecules quickly and at low cost. [0003] 17b-estradiol is considered to be the most potent and potentially influential class of endocrine disruptors, widely present in rivers, soil, atmosphere and agricultural products. It is mainly derived from pesticides, exhaust gas, food additives, human and animal excrement and domestic sewage. Even at extremely low concentrations, it will have a significant impact on organisms. Its content level is comparable to that of prostate cancer, breast cancer, ovarian cancer, There is a significant association between uterine cancer and male reproductive disorders. Traditional 17b-estradiol detection methods mainly include hig...

AI Technical Summary

Problems solved by technology

However, the detection sensitivity of the colorimetric method based on G tetramer is low, and the sensitivity needs to be further improved.

The traditional signal amplification technology is mainly PCR and a series of signal amplification technologies relying on tool enzymes, but the use of enzymes not only increases the cost and operation steps, but also the storage of enzymes is more troublesome, so it is necessary to develop a tool-free enzyme signal amplification technology. technology to improve detection sensitivity

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