Probe applied to real-time detection of nucleic acid and application thereof

Abstract

The invention discloses a probe applied to real-time detection of nucleic acid and application thereof, wherein the probe is an oligonucleotide which comprises 10 partial regions; the first region (1) at the 3'end and the eighth region (8) at the 5'end form a first double chain, so that a fluorescent group or a fluorescence donor (10) connected by the 5'end is close to a quencher or a fluorescence receptor (9) connected by the 3' end, and no fluorescence is emitted; the second region (2) and the fourth region (4) are paired, and the fifth region (5) and the seventh region (7) are paired, so as to form the other two sections of double chain; the third region (3) and the sixth region (6) form two annular structures respectively; the basic groups and target sequences of the third region (3), the fourth region (4), the fifth region (5) and the sixth region (6) of the probe are complementary to form a target sequence combination region. The invention further discloses application of the probe on a real-time PCR (Polymerase Chain Reaction) detection method of the target sequence. The probe is strong in specificity and low in fluorescent background. In addition, the probe has the reverse complementary double-stranded structure, so that the specificity of the probe can be adjusted by adjusting the length of the complementary regions during designing.

Description

technical field [0001] The invention relates to the field of molecular detection, in particular to a novel probe for real-time fluorescence detection of nucleic acid. The invention also relates to the technical application of the "pin shape" probe. Background technique [0002] Polymerase chain reaction (PCR) technology is a simple, fast, highly sensitive and specific gene amplification method, and its process usually includes 20-50 cycles consisting of three steps of denaturation, annealing and extension. It can specifically amplify the target nucleic acid sequence to several million times within 1.5-3 hours. In recent years, due to the appearance of real-time PCR instrument, gel electrophoresis analysis is no longer required for ordinary PCR, so post-PCR operations are not necessary, and PCR pollution is eliminated. Therefore, real-time PCR technology is more and more widely used in clinical practice. [0003] Real-time fluorescent quantitative PCR detection technology ...

AI Technical Summary

Problems solved by technology

However, as mentioned above, the probes generally have complex design, high fluorescence background, and limited ability to recognize single-base mutations.

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