Novel method for detecting and quantifying mononucleotide variation

Abstract

A novel method for detecting and quantifying mononucleotide variation is characterized in that a probe is combined with locked nucleic acid (LNA) modification to improve the difference between Tm1 and Tm2, the Tm1 is the Tm value when the probe is combined with a completely matched sequence, the Tm2 is the Tm value when the probe is matched with a mononucleotide variation sequence, and the modified probe is used for distinguishing and quantifying mononucleotide variation sites. The nucleotide in the probe is subjected to LNA locked nucleic acid modification, and the temperature difference between Tm1 and Tm2 is improved, so that the distinguishing capability of the probe on a single nucleotide variation sequence is improved, and a template with lower mutation content can be detected.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a new method for detecting and quantifying single nucleotide variation. Background technique [0002] The identification of single nucleotide variation is of great significance in drug monitoring and other aspects. For example, a specific gene mutation is an important biomarker for tumor-targeted therapy, and gene mutation detection can detect drug-resistant mutations as early as possible before the disease progresses, helping doctors make clinical treatment decisions and strive for treatment opportunities for patients. [0003] At present, the detection of single nucleotide variation mainly uses a real-time fluorescence quantitative PCR (polymerase chain reaction, polymerase chain reaction) platform. The real-time fluorescent quantitative PCR platform is a method of measuring the total amount of products after each cycle of polymerase chain reaction (PCR) with fluorescent chemicals...

AI Technical Summary

Problems solved by technology

[0005] At present, ARMS technology has the following disadvantages: 1. ARMS technology is limited by the sequence at the end of the primer, and the design is relatively limited; 2. ARMS primers need to strictly control the reaction solution ratio and reaction temperature to meet the required detection requirements, and the reproducibility is low. Poor; 3. The specificity of ARMS primers is limited. Specificity refers to the concentration of non-detectable nucleotide sites that can be tolerated. The higher the concentration that can be tolerated, the lower the proportion of mutations that can be detected, which is more conducive to low mutations Accurate detection of content samples

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