Nucleic acid detection method applying multiplex PCR array technology

Abstract

The present invention discloses a nucleic acid detection method applying a multiplex PCR array technology, and further relates to universal tag (UT) sequences, a primer pair comprising the universal tag sequences and specific detection primers (5'-UT primer+specific detection primer-3'), a kit containing the universal tag sequences or the primer pair, and applications of the sequences, the primer pair and the kit in nucleic acid biological detection, wherein the UT sequences meet the following conditions: 1) the UT sequences of the upstream primer and the downstream primer are different, and Tm value difference does not exceed +-1 DEG C, and is about 5-10 DEG C greater than a detection primer Tm value; and 2) the UT sequences are the randomly-designed DNA fragments, and do not have homology with the sequence of the organism requiring detection. According to the detection method, sensitivity and specificity of nucleic acid detection are increased through combination of the mPCR and the universal tag sequences.

Description

1. Technical field [0001] The invention relates to a nucleic acid detection method, in particular to a nucleic acid detection method using multiple PCR array technology, and belongs to the field of biotechnology. 2. Background technology [0002] Multiplex PCR technology (multiplex PCR, mPCR) is another major innovation in this technical field since the birth of PCR technology in the 1980s. Traditional PCR technology only uses a pair of primers to generate a nucleic acid fragment through amplification, which is mostly used for the identification of a single target sequence. However, mPCR is a PCR reaction in which more than two pairs of primers are used in the same PCR reaction system to simultaneously amplify multiple nucleic acid fragments. The reaction principle, reaction reagents and operation process are the same as those of classic PCR. [0003] The biggest feature of mPCR is that it improves the efficiency of detection, making PCR a high-throughput method: (1) high e...

AI Technical Summary

Problems solved by technology

Simultaneous PCR detection of multiple gene target sequences often leads to the phenomenon that in order to ensure the specificity of the primers, it is difficult to unify the Tm values ​​of the primers with different lengths and GC content, which affects the annealing temperature of the entire mPCR system. Difficulty with program setup, which reduces the sensitivity of mPCR detection

Images