Method for detecting related gene mutations of diabetes drug treatment as well as special chip and kit thereof

Abstract

The invention discloses a method for detecting related gene mutations of diabetes drug treatment as well as a special chip and a kit thereof. The method comprises the following steps: taking a genome to be detected from a human tissue as a template, carrying out multiple PCR amplification by a primer group that is designed aiming at special mutant sites and DNA polymerase without 3'-5' end exonclease activity, then hybridizing the obtained PCR product and an allele specific probe on the gene chip, and confirming mutation types of related genes of diabetes drug treatment according to the hybridizing result. The invention can detect known gene mutations closely related to the individual difference of anti-tumor medicine reaction in comprehensive, systemic and high-flux ways and greatly improve the accuracy and the specificity of chip detection.

Description

technical field [0001] The invention relates to a method for identifying gene mutation types related to diabetes drug treatment in the field of gene analysis, as well as a dedicated chip and a kit. Background technique [0002] Diabetes, as a major disease that plagues human physical and mental health, has become a global public health problem. According to the World Health Organization, there will be 380 million diabetic patients in the world in 2025. In my country, the incidence of diabetes has reached 2%, and the number of diagnosed diabetic patients has reached 40 million. The total number of patients has ranked second in the world, and is increasing at a rate of 1 million per year. For diagnosed diabetic patients, the most effective treatment is drug hypoglycemic therapy. [0003] The commonly used diabetes treatment drugs in clinical practice are shown in the following table: [0004] Table 1 [0005] category representative drug Sulfonylur...

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Problems solved by technology

[0015] Although the above-mentioned chip technology can theoretically use the phenomenon of poor stability of hybrids formed by probes containing mismatched bases and target genes to detect base changes in gene sequences, it is often due to different forms of base mismatches, gene The actual situation is much more complicated than the theoretical situation due to factors such as different sequences and differences in the high-level structure of the gene. In order to improve the detection accuracy of the probe for a single base change, it is generally preferred to design the probe to be shorter. The specificity of gene recognition and the maintenance of high hybridization signal intensity cannot make the probe too short, and even the probes of the same length will have different GC content and the distance of the detected mutation base on the probe. The relative position difference makes this probe-based hybridization process very complicated, and the hybridization specificity between the short probe and the target sequence becomes very prominent at this time. The specific performance is that the wild-type probe and the mutant probe Non-specific hybridization signals will appear during the hybridization of needles, and it is impossible to achieve "all or none" of detection signals. Only by strictly controlling the hybridization conditions and repeatedly exploring can the genotypes be accurately distinguished

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