Targeting medicament curative effect related gene mRNA expressive level detection Luminex and detection method

Abstract

The invention discloses a receptor tyrosine kinases inhibiting type targeting medicament curative effect related gene mRNA expressive level detection Luminex. The detection Luminex mainly comprises a micro-ball, supporting extending probes, and amplification extension probes. The micro-ball is coupled with a supporting probe modified by amino; the supporting extending probes are used for connecting the supporting probe and a target gene mRNA, and each supporting extending probe comprises a specific sequence the terminal 5' of which can combine with a corresponding target gene mRNA, a spacer arm sequence, and a sequence the terminal 3' of which can complementarily pair with the corresponding specific sequence of the supporting probe at; and the amplification extension probes are used for connecting the target gene mRNA and information detection components, and each amplification extension probe comprises a specific sequence the terminal 5' of which can combine with the corresponding target gene mRNA, a spacer arm sequence, and a sequence the terminal 3' of which can complementarily pair with a markup probe. The invention also discloses a detection method of the receptor tyrosine kinases inhibiting type targeting medicament curative effect related gene mRNA expressive level by using the liminex. In the invention, the process can be carried out without the steps of RNA extraction, reverse transcription, PCR and the like and the detection result is influenced less by the quality of RNA in the sample, thereby the accuracy of the detection result is guaranteed; on the other hand, because the liminex technology enables a parallel multi-index detection to be realized, multiple inside genes can be set in one detection, thereby the detection result is more reliable.

Description

Technical field [0001] The invention belongs to the field of molecular biology, and relates to medicine and biotechnology, and specifically relates to a liquid chip and a detection method for detecting the mRNA expression level of genes related to the therapeutic effect of receptor tyrosine kinase inhibitor targeted drugs. Background technique [0002] Tyrosine kinase occupies a very important position in the intracellular signal transduction pathway, regulating a series of physiological processes such as cell growth, differentiation, and death, and its abnormal expression will lead to the disorder of cell proliferation regulation, which will lead to tumorigenesis. In addition, the abnormal expression of tyrosine kinase is also closely related to tumor invasion and metastasis, tumor angiogenesis, and tumor resistance to chemotherapy. Drug development with tyrosine kinase as the target has become an international anti-tumor drug research Hot spot. [0003] Tyrosine kinases include ...

AI Technical Summary

Problems solved by technology

Among them, RISH is mainly used to analyze the distribution of RNA in tissues or cells to understand the expression of specific genes, and the process is long and cumbersome, so it is not suitable for clinical applications; while reverse transcription quantitative PCR method and real-time fluorescent quantitative PCR method, First of all, both methods require RNA extraction and reverse transcription, and the detection results are greatly affected by RNA degradation; second, these two methods only use a pair of primers for PCR amplification, which is prone to non-specific binding, resulting in false positives. The positive rate is high; again, these two methods generally only have one housekeeping gene as a control, and their accuracy is easily affected by pathological conditions; defect

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