Kit, primer, probe and detection agent for detecting tumor gene variation

Abstract

The invention discloses a kit, a primer, a probe and a detection agent for detecting tumor gene variation. The kit, the primer, the probe and the detection agent for detecting the tumor gene variationcan be used for detecting tumor driving gene EGFR mutation, KRAS mutation, BRAF mutation, HER2 mutation, MET mutation, EML4-ALK fusion and ROS1 fusion. The fluorescent probe for detecting tumor genevariation, disclosed by the invention, has the advantages that the fluorescent probe is specifically combined with a mutant type DNA template in the PCR process but not combined with a wild type DNA template, so that a mutant type amplification result generates a fluorescence signal, and a wild type amplification result does not generate the fluorescence signal; a final judging result is that themutant type amplification result has a Ct value and the wild type amplification result has no Ct value. The primer and the probe for detecting tumor gene variation, disclosed by the invention, have the advantages of high specificity, simple kit operation, high detection sensitivity and objective and accurate qualitation of tumor gene variation.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit, primers, probes and detection agents for detecting tumor gene variation. Background technique [0002] Epidermal growth factor receptor (EGFR) is a multifunctional glycoprotein widely distributed in the cell membranes of various tissues in the human body. It is one of the four members of the HER / ErbB family, so it is also known as HER1 or ErbB-1 . The gene encoding human EGFR is located in the p13-q22 region of chromosome 7, with a total length of 200kb, consisting of 28 exons, encoding 1186 amino acids, and its glycoprotein molecular weight is about 170kDa, which is related to HER2 / ErbB-2 / Neu / p185, HER3 / ErbB-3, HER4 / ErbB-4, etc. are classified into the HER / ErbB family and belong to receptor tyrosine kinases (receptor tyrosinekinase, RTK). In patients with non-small cell lung cancer, the application of targeted therapeutic drugs targeting EGFR gene has achieved r...

AI Technical Summary

Problems solved by technology

The traditional ARMS-PCR detection principle is to judge negative and positive by comparing the Ct difference between the wild-type and mutant amplification curves, and the Ct value is determined by artificially adjusting the baseline (Threshold). However, the wild-type and mutant amplification curves often are not parallel, different baseline adjustment values ​​will lead to different Ct differences

Therefore, the interpretation of the results of the ARMSPCR detection method is too subjective, and it is easy to cause human misjudgment, especially for weakly positive samples, where the possibility of misjudgment is extremely high

In addition, the traditional ARMS PCR cannot avoid the misjudgment of the results caused by the external control and the change of the ratio of sample DNA in genomic DNA caused by the difference of nucleic acid extraction.

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