Multi-gene detection kit for guiding administration of 5-fluorouracil and detection method of multi-gene detection kit

Abstract

The invention discloses a multi-gene detection kit for guiding administration of 5-fluorouracil and a detection method of the multi-gene detection kit. The multi-gene detection kit comprises ultrapure water, an X solution, a 10*PCR (Polymerase Chain Reaction) buffer, a PCR primer, a 25mM magnesium chloride solution, a DNA (Deoxyribonucleic Acid) polymerase and a positive control, and is characterized in that the PCR primer comprises six forward and reverse amplification primers with different genotypes on seven SNP (Single Nucleotide Polymorphism) sites of genes related to the administration of 5-fluorouracil, and forward and reverse amplification primers of a DNA internal reference and a reaction internal reference, and gene sequences of the forward and reverse amplification primers are shown as SEQ ID NO. 1-32 (Sequence Identifier Numbers 1-32). The detection method of the multi-gene detection kit comprises the steps of collecting a sample, extracting nucleic acid, conducting a PCR by taking the extracted nucleic acid as a template, and conducting capillary electrophoretic separation on the sample with a GeXP genetic analyser. The multi-gene detection kit and the detection method have the advantages of strong specificity, high accuracy, flux and reliability, low cost and no false negative result.

Description

technical field [0001] The invention relates to a multiple gene detection kit and a detection method thereof, in particular to a multiple gene detection kit for guiding the administration of 5-fluorouracil and a detection method thereof. Background technique [0002] 5-FU drugs belong to the fluorouracil class of antimetabolite chemotherapy drugs. These drugs are converted into 5-FU in the body to inhibit deoxythymidylate synthase, thereby affecting the synthesis of DNA and achieving the purpose of inhibiting tumor growth. Therefore, the use of such drugs will inevitably damage the normal cell division in the patient's body, resulting in many side effects of chemotherapy. 5-FU drugs have a wide anticancer spectrum and are one of the most widely used anticancer drugs. However, in the clinical application of 5-FU drugs, there are problems of low treatment efficiency and large side effects, and there are significant individual differences, some patients benefit, and some patie...

AI Technical Summary

Problems solved by technology

Its disadvantages are (1) low throughput: it is not suitable for the detection of multiple SNP sites; it is difficult to set internal control genes

(2) High cost: the cost of probe labeling is high; if you need to obtain all relevant SNP information, you need to conduct multiple detection tests, and the superimposed cost is more expensive

Disadvantages: 1) The hybridization kinetics between different SNP sites are different, and the conditions are difficult to control when performing simultaneous detection of multiple sites; 2) The technology is expensive and complicated: each sample requires a chip, and the cost is more than ￥1000 / sample , which is not conducive to large-scale promotion; the synthesis and fixation of probes are relatively complicated, especially the production of high-density probe arrays, which is the main speed-limiting step; 3) poor repeatability, low accuracy, and false positive and false negative results are prone to occur; 4) Low sensitivity: the chip method requires a large amount of nucleic acid, and generally requires multiple PCR amplification first. Due to the large number of primers, it is easy to produce dimers and hairpin structures by itself, or the purpose of amplification is caused by different Tm values. Different fragment efficiencies will affect the detection sensitivity; 5) Due to the variety of chips, it is difficult to formulate a unified quality control standard

[0014] At present, there are no relevant research reports on multiple gene detection kits and detection methods based on GeXP multiple gene expression genetic analysis system to guide 5-fluorouracil medication at home and abroad

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