Incision enzyme mediated real-time multiple cross nucleic acid displacement amplification technology and application

Abstract

The invention discloses an incision enzyme mediated real-time multiple cross nucleic acid displacement amplification technology and an application. The invention provides a pair of cross primers which contain restriction incision enzyme specific recognition sequences at an end 5', and are marked with fluorescent groups and quenching groups, a pair of displacement primers and 3-6 amplification primer constant-temperature amplification target gene segments; and the incision enzyme mediated real-time multiple cross nucleic acid displacement amplification technology can carry out visible detection, electrophoresis detection, real-time turbidity detection and real-time fluorescence detection. The method is convenient, quick, sensitive, specific and suitable for detecting various nucleotide fragments.

Description

technical field [0001] The invention discloses a nucleic acid amplification technology, which belongs to the field of molecular biology. Background technique [0002] In the field of modern medicine and biology, nucleic acid amplification is an indispensable technology, widely used in clinical testing, basic research, archaeological research, epidemiological research, transgenic research and other fields. Among the nucleic acid amplification technologies that have been developed and designed, PCR amplification technology is the first established in vitro nucleic acid amplification technology, which has epoch-making significance. This technology has been widely used in medical, biological and chemical related fields. [0003] When using PCR technology for nucleic acid amplification, it relies on complex and expensive thermal cycle equipment, and the detection of amplification products is relatively complicated, requiring a complex process and equipment. Therefore, the applic...

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Problems solved by technology

[0008] At present, the detection of Shigella and Salmonella mainly relies on traditional enrichment culture and biochemical identification. This method takes about 5 to 7 days, including enrichment, selective culture and subsequent biochemical identification. Its disadvantage is time-consuming and labor-intensive. , the interpretation of biochemical results depends on human subjective judgment, resulting in poor repeatability of results and easy misjudgment

With the rapid development of nucleic acid diagnostic technology, some PCR-based diagnostic technologies (such as ordinary PCR technology, fluorescent PCR technology) are used for the rapid detection of Shigella and Salmonella, but these methods rely on expensive equipment, Requires subsequent electrophoresis, expensive probe synthesis, and skilled operators

It cannot be carried out for some backward laboratories, thus limiting the application of these technologies

At present, the PCR method and real-time PCR method using these detection technologies to detect Shigella and Salmonella have poor diagnostic sensitivity and a long detection process, which is not conducive to rapid detection and emergency detection.

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