Amplification primer, detection probe and liquid phase chip for EGFR gene mutation detection

Abstract

The invention belongs to the technology of medical IVD (in vitro diagnosis) and particularly relates to an amplification primer, a detection probe and a liquid phase chip for EGFR (epidermal growth factor receptor) gene mutation detection. The liquid phase chip for EGFR gene mutation detection comprises PCR (polymerase chain reaction) amplification primers, LDR (ligase detection reaction) connection probes and Anti-Tag sequence coated microspheres, wherein the amplification probes comprise four pairs of primers for exons 18, 19, 20 and 21; and the connection probes comprise eight pairs of connection probes for eight primary mutation sites related with drug therapeutic effect of a tyrosine kinase inhibitor (TKI). The liquid phase chip can be used for carrying out joint detection on mutation of eight EGFR genes and accurately analyzing the eight mutation sites at the same time by single reaction, and has the characteristics of low price and suitability for parallel detection of a large amount of samples.

Description

technical field [0001] The invention belongs to medical in vitro diagnostic technology, and specifically relates to an amplification primer, a detection probe and a liquid phase chip for detecting mutations on the EGFR gene related to the curative effect of tyrosine kinase inhibitor (TKI) drugs. Background technique [0002] Epidermal growth factor receptor (EGFR) is a member of the cell surface receptor tyrosine kinase HER / ErbB family, which can specifically bind to ligands and make itself Phosphorylation, thereby regulating cell growth or survival. [0003] Molecular targeted therapy targeting EGFR is becoming increasingly prominent in the treatment of NSCLC, among which EGFR tyrosine kinase inhibitors (TKI), such as gefitinib and erlotinib, are the most important. Pre-clinical trials of NSCLC have shown that after failure of platinum-based drugs combined with chemotherapy, gefitinib still has an objective response rate of 12% to 18%, a disease control rate of >40%, an...

AI Technical Summary

Problems solved by technology

DNA sequencing is currently the gold standard for detecting EGFR gene mutations by comparing forward and reverse bidirectional sequencing. However, this method requires amplification, purification, and sequence analysis of sequencing samples. There are many steps, and the reaction takes a long time, which may increase the probability of errors.

In addition, the DNA sequencing method also has many disadvantages such as cumbersome operation, complex interpretation of results, and high requirements for the environment and operators. Therefore, it is only suitable for a few well-equipped laboratories, and few hospitals at all levels can carry out it independently.

Although the real-time fluorescence quantitative PCR method can detect more samples in a short period of time, each reaction can only detect one or a few mutations, which cannot achieve high throughput, and there is also the possibility of false negatives

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