Fluorescent probe for fluorescent quantitative PCR reactions

Abstract

The invention relates to a fluorescent probe for fluorescent quantitative PCR reactions. The fluorescent probe is characterized in that the probe comprises an oligonucleotide sequence, the 5' terminus is a specific oligonucleotide sequence with an amplified target sequence and modifies the fluorescence groups; the 3' terminus is a nucleotide sequence, forms a reversely-complimentary palindrome structure with the 5' terminus, and modifies the fluorescence quenching groups. Under certain conditions, the probe can form an intramolecular secondary composition, and the fluorescent groups get close to the quenching groups so as to quench the fluorescence signals released by the fluorescence groups. Compared to the common fluorescent probe, the provided fluorescent probe has the advantages of high specificity, high sensitivity, and low fluorescent quenching efficiency, thus the signal to noise ratio (SNR) of the reactions is higher, and the provided fluorescent probe can be applied to realtime fluorescence quantitative PCR detection reactions and all end point fluorescence detection reactions.

Description

technical field [0001] The present invention relates to an oligonucleotide fluorescent probe, more specifically to a fluorescent probe for fluorescent quantitative PCR reaction, the probe can be applied to real-time fluorescent quantitative PCR reaction detection, etc., belongs to biotechnology field. Background technique [0002] Fluorescence Quantitative Polymerase Chain Reaction (FQ-PCR, qPCR), as a new generation of molecular biology research tools, has become a mainstream technology platform for molecular diagnosis, clinical testing and basic biomedical research, and is widely used in infectious diseases, tumors, Molecular diagnosis of genetic diseases, cardiovascular and other diseases. [0003] Early real-time PCR nucleic acid detection technologies used fluorescent dyes to indicate the accumulation process of PCR reaction products, such as SYBR GREEN, EVE GREEN, etc. The dye method is simple, low in cost, and does not require the design of special probes. However, ...

AI Technical Summary

Problems solved by technology

The TaqMAN probe that specifically binds to the target molecule will excise the fluorescent molecule at the 5' end through the action of the 5' exonuclease of Taq DNA polymerase during the PCR amplification process to release the fluorescent signal, and the destroyed probe will The needle cannot be reversibly restored, so although the TaqMAN probe has greater signal amplification and higher detection sensitivity; however, due to the long distance between the fluorescent group and the quencher group, the fluorescence quenching is incomplete and the fluorescence background is high

In contrast to molecular beacons, the fluorescent molecules are adjacent to the quencher molecules, the distance is relatively close, and the fluorescence quenching is complete, so the fluorescence background is low; however, the molecular beacons are not destroyed during the PCR reaction, so the signal amplification only depends on the PCR product Cumulative, less sensitive detection

Scorpion probes are similar to molecular beacons, but their disadvantage is that the signal amplification ability is inferior to that of TaqMan probes

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