Method for isothermal amplification of nucleic acids and method for detecting nucleic acids using simultaneous isothermal amplification of nucleic acids

Abstract

The present invention relates to a method for isothermal amplification of nucleic acids and a method for detecting nucleic acids, which comprises characterized in simultaneous isothermal amplification of nucleic acids and a signal probe to a method for isothermal amplification of target nucleic acids using an external primer set and RNA / DNA hybrid primer set, and a method for detecting amplification products by amplifying nucleic acids and a signal probe simultaneously using an external primer set, RNA-DNA hybrid primer set and DNA-RNA-DNA hybrid probe. The method according to the present invention is convenient compared with the conventional method, it is possible to amplify the target nucleic acids rapidly and exactly without a risk of contamination, and it can simultaneously amplify a signal probe, so that it can be applied to various genome project, such as detection and identification of a pathogen, detection of gene modification causing predetermined phenotype, detection of hereditary diseases or determination of sensibility to diseases, estimation of gene expression and apply to genome project, thus being useful for molecular biological studies and disease diagnosis.

Description

technical field [0001] [01] The present invention relates to a nucleic acid isothermal amplification method and a nucleic acid detection method comprising simultaneous nucleic acid and signal probe isothermal amplification for detection of amplified products, more specifically, to a method using external primers Target nucleic acid isothermal amplification method for pair and RNA / DNA hybridization primer pairs, and a method for simultaneously amplifying nucleic acid and signaling probe by using external primer pairs, RNA-DNA hybridization primer pairs, and DNA-RNA-DNA hybridization probes Amplification product detection method. Background technique [0002] [02] Nucleic acid amplification techniques are very useful for detecting and analyzing small amounts of nucleic acids. The high sensitivity of nucleic acid amplification to target nucleic acids has led to the development of techniques for the detection of specific nucleic acids based on genetic isolation, which can be us...

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Problems solved by technology

Therefore, PCR technology has the following disadvantages: its cost is very high; it has relatively low specificity; it requires extreme performance criteria to re-perform the results

[0016] The problem with [16] is that amplification methods using thermal cycling processes such as PCR require heating of the block to reach the "target" temperature for each cycle, and require a delay time before the heating block reaches the target temperature, thus taking a long time. Long time to complete the amplification reaction

[0022] [22] However, the disadvantage of the CPT method is that its amplification efficiency is 103-106, which is relatively low, so it is difficult to use it alone in diagnosis, and its usage is also complicated because the signal probe is passed through such as Traditional nucleic acid amplification such as PCR is used to increase the number of special regions of the target nucleic acid and then amplify separately, and it requires high cost and a long time

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