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Kit for synchronously detecting related gene expression level of 14 antitumor drugs by using paraffin embedding biopsy sample, and detection method thereof

A gene expression level, paraffin-embedded technology, used in biochemical equipment and methods, measuring devices, and microbial determination/inspection, etc. and other problems, to achieve the effect of saving detection cost, superior low cost, and solving easy pollution

Active Publication Date: 2013-09-18
NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there are also the following disadvantages: 1) High cost: each sample needs 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 more complicated, especially the production of high-density probe arrays , is the main rate-limiting step; 2) cannot be accurately quantified, and the repeatability is poor; 3) the sensitivity is low and the amount of nucleic acid required is large, and because there are many types of chips, it is difficult to formulate a unified quality control standard
[0016] At present, there are no relevant research reports on the kits and detection methods for the simultaneous detection of the expression levels of 14 anti-tumor drug-related genes based on the GeXP multiple gene expression genetic analysis system at home and abroad.

Method used

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  • Kit for synchronously detecting related gene expression level of 14 antitumor drugs by using paraffin embedding biopsy sample, and detection method thereof
  • Kit for synchronously detecting related gene expression level of 14 antitumor drugs by using paraffin embedding biopsy sample, and detection method thereof
  • Kit for synchronously detecting related gene expression level of 14 antitumor drugs by using paraffin embedding biopsy sample, and detection method thereof

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Experimental program
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specific Embodiment 2

[0070] The present invention is a method for synchronous detection of 14 anti-tumor drug-related genes using paraffin-embedded section samples, collecting patient tumor tissue to extract nucleic acid, performing reverse transcription and PCR reaction with patient nucleic acid as a template, and finally separating samples by capillary electrophoresis ,Specific steps are as follows:

[0071] 1. Production of a kit for synchronous detection of 14 anti-tumor drug-related genes based on the GeXP multiple gene expression genetic analysis system using paraffin-embedded section samples. The components included in the kit are the same as those in Example 1 above;

[0072] 2. Collect samples and extract nucleic acids

[0073] Collect isolated cultures of paraffin-embedded section samples of tumor tissue from patients, and extract nucleic acids from the isolated cultures;

[0074] 3. Reverse transcription (RT) reaction using patient nucleic acid as a template

[0075] 1) Add reagents ...

specific Embodiment 3

[0099] Detection Kit Sensitivity and Specificity Analysis

[0100] Sensitivity analysis: After diluting the positive control substance by a certain ng value, it is detected by PCR amplification and capillary electrophoresis until no signal is detected. The ng value is the lowest detection line, which is the sensitivity of the kit. The sensitivity of this kit is 1.0ng.

[0101] Specificity analysis: Single-plex PCR amplification is detected as a single peak of the target fragment size by capillary electrophoresis.

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Abstract

The invention discloses a kit for synchronously detecting related gene expression level of 14 antitumor drugs by using a paraffin embedding biopsy sample, and a detection method thereof. The kit comprises DEPC (diethylpyrocarbonate) water, 5*RT buffer solution, a reverse transcription primer, a reverse transcription enzyme, an X solution, 10*PCR buffer solution, a PCR primer, 25mM magnesium chloride solution, DNA (deoxyribonucleic acid) polymerase and a positive reference substance; the reverse transcription primer comprises related genes of the 14 antitumor drugs, and RT amplification primers of an RNA (ribonucleic acid) internal reference; gene sequences are shown in SEQ ID NO.1 to NO.18; the PCR primer includes related genes of the 14 antitumor drugs, and positive and negative PCR amplification primers of a DNA internal reference; the gene sequences are shown in SEQ ID NO.19 to NO.38. The kit has the advantages of strong specificity, high sensitivity, high flux, strong reliability, low cost and absence of false-negative result.

Description

technical field [0001] The present invention relates to a kit for gene expression levels related to anti-tumor medications and a detection method thereof, in particular to a reagent for synchronously detecting the expression levels of 14 anti-tumor medication-related genes using a paraffin-embedded section sample based on a GeXP multiple gene expression genetic analysis system box and its detection method. Background technique [0002] In recent years, pharmacogenetics / pharmacogenomics has made breakthroughs in the research on the mechanism of action of anti-tumor drugs, and found that the killing effect of certain anti-tumor drugs on tumor cells is related to a specific gene (or group) Expression and / or polymorphisms of . Through the detection of related genes, predicting the efficacy of chemotherapy and targeted drugs, and selecting appropriate drugs for individualized chemotherapy has become a reasonable choice to improve efficacy and reduce ineffective treatment. [00...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N27/447
Inventor 南丽徐冰吴勇
Owner NINGBO HEALTH GENE TECHNOLOGIES CO LTD
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