Constant temperature synchronous amplification detecting process for nucleic acid and use thereof

Abstract

The invention discloses a constant temperature synchronous amplified detection method of nucleic acid as well as an application thereof. The method comprises the following steps of: 1) mixing reactants containing the following components: a. a nucleic acid sample under test; b. a primer 1: a 3`-terminal of the primer can be hybridized at a 3`-terminal or near the 3`-terminal of the nucleic acid sample under test, and a 5`-terminal is a promotor sequence; c. a primer 2: the primer can be hybridized with a 5`-terminal of a negative strand of the nucleic acid sample under test; d. one or a plurality of fluorescent probes; e. at least one DNA polymerase relied by RNA; and f. at least one RNA polymerase that can identify the promotor sequence; and 2) carrying out the constant temperature amplification reaction to the mixed reactants in a closed vessel, detecting the changes of fluorescence signals in a reaction system synchronously by a detector, and conducting the quantitative and qualitative detection to the nucleic acid sample according to the time and intensity of fluorescence signal changing. The method and the application have the advantages of low pollution, constant temperaturein the reaction process, high detection sensitivity, fast detection speed, low requirements on equipment and instrument and low cost, and are applied to nucleic acid testing in fields such as clinicalexamination and blood screening.

Description

technical field [0001] The invention relates to a biological detection technology, in particular to a detection method for synchronously performing constant temperature amplification and detection of nucleic acid, and its application in nucleic acid detection in clinical testing, blood screening, food safety inspection and environmental monitoring. Background technique [0002] In the process of nucleic acid detection, in most cases, the content of nucleic acid in the sample is low. In order to achieve a certain sensitivity and accuracy in the detection, it is necessary to amplify the nucleic acid fragments to be detected in the sample. In 1985, the emergence of Polymerase Chain Reaction (PCR) technology promoted the development of nucleic acid detection technology, which has become an important means in nucleic acid detection technology. Subsequently, derivatives such as reverse transcriptase-polymerase chain reaction (Reverse Transcriptase-PCR, RT-PCR for short), real-time...

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Problems solved by technology

At present, in the field of nucleic acid diagnosis, there are no related technical reports or inventions that simultaneously perform isothermal amplification and detection.

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