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Genetic evaluation and suit method for individualized tumor therapy

A gene and tumor technology, applied in the field of tumor personalized therapy gene evaluation and kits, can solve the problems of toxic side effects, short survival period, affecting the efficacy of platinum drugs, and achieve the effect of avoiding toxic side effects and improving curative effect.

Inactive Publication Date: 2012-10-17
SHANG OUTDO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

On the one hand, DPD functional defects can increase the concentration of 5-FU active metabolite 5-fluorouridine deoxynucleoside, but at the same time, it can also produce serious and even fatal toxic side effects
XRCC1 Arg399Gln affects the efficacy of platinum-based drugs. Compared with homozygous 399Arg patients, patients with 399Gln are less effective in oxaliplatin / 5-FU therapy and have shorter survival time (Liu B, Wei J, Zou Z, Qian X, Nakamura T, Zhang W, Ding Y, Feng J, Yu L. Polymorphism of XRCC1 predicts overall survival of gastric cancer patients receiving oxaliplatin-based chemotherapy in Chinese population. Eur J Hum Genet 2007 Oct; 15(10): 1049-1053. Stoehlmacher J, Ghaderi V, Iobal S, Groshen S, Tsao-Wei D, Park D, Lenz HJ. A polymorphism of the XRCC1 gene predicts for response to platinum based treatment in advanced colorectal cancer. Anticancer Res 2001 Jul-Aug; 21(4B ): 3075-3079. Martinez-Balibrea E, Manzano JL, Martinez-Cardus A, Moran T, Cirauqui B, Catot S, Taron M, Abad A. Combined analysis of genetic polymorphisms in thymidylate synthase, uridine diphosphate glucoronosyltransferase and X-ray cross complementing factor 1 genes as a prognostic factor in advanced colorectal cancer patients treated with 5-fluorouracil plus oxaliplatin or irinotecan. Oncol Rep 2007Mar;17(3):637-645.)

Method used

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  • Genetic evaluation and suit method for individualized tumor therapy
  • Genetic evaluation and suit method for individualized tumor therapy
  • Genetic evaluation and suit method for individualized tumor therapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: the extraction of sample DNA

specific Embodiment approach

[0033] Embodiment 1: the extraction of sample DNA

[0034] Genomic DNA in human peripheral blood was extracted using the FlexiGene DNA Kit (QIAGEN, Cat. No. 51206).

[0035] The specific method is: add 750 μl Buffer FG1 to 300 μl blood sample, invert it up and down 5 times to mix well. Then centrifuge at 12,000 rpm for 1 min. After centrifugation, discard the supernatant, then add 150 μl Buffer FG2 and 1.5 μl protease solution, shake immediately until the precipitate is completely dissolved. Next, centrifuge for 3-5 seconds, and then bathe in water at 65°C for 5 minutes. When the solution turns from red to olive green, add 150 μl of 100% isopropanol, invert the centrifuge tube up and down fully, and mix it until the DNA precipitates in the form of linear or lumps visible to the naked eye. Then centrifuge at 12,000 rpm for 3 min. After centrifugation, discard the supernatant, then add 150 μl 70% ethanol, and shake for 5 seconds. Then centrifuge again at 12,000 rpm for 3 mi...

Embodiment 2

[0037] Example 2: Detection of SNP sites by chip hybridization

[0038] 1. Preparation of gene chip

[0039] (1) Probe dissolution

[0040] Dilute each probe of the sequence shown in SEQ ID NO: 27 to SEQ ID NO: 58 with TE solution, and the final concentration is 10 mM. Mix the probe with a concentration of 10mM and the PBS solution with a concentration of 200mM in a medium volume of a 384-well plate, seal the 384-well plate with an adhesive sheet, shake at room temperature for 2 minutes, centrifuge, and store at -20°C for sample application .

[0041] Table 1 Probes for different SNP sites

[0042]

[0043] (2) Spotting

[0044] The pre-designed and synthesized probes are loaded onto the solid-phase carrier substrates made of glass slides, silicon wafers, etc. through contact spotting or inkjet spotting. The film base adopts Cell Associates CSS-100 aldehyde base film base, and the Ominigrid 100 model spotting instrument of GeneMachine Company is applied at a humidity o...

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Abstract

The invention discloses a genetic evaluation and suit method for individualized tumor therapy. The genetic evaluation and package method includes steps of: 1) extracting a DNA sample of a subject; 2) carrying out target gene PCR amplification, purification, fragmentation and fluorescein labeling on the sample DNA; 3) carrying out gene chip hybridization on fluorescein labeled PCR products to detect SNP sites of the target gene; and 4) analyzing curative effect and toxicity of antitumor drug on the subject, according to the genetic typing results obtained in the step 3). Therapeutic effect evaluation of tumor patient can be conducted through the related genetic typing, thus formulating the most scientific individualized therapy plan to improve efficacy, reduce or even avoid the toxic and side effects.

Description

technical field [0001] The present invention relates to a tumor individualized therapy gene evaluation and kit method, in particular to a chemotherapy efficacy gene evaluation and kit method. Background technique [0002] With the development of my country's social economy and changes in people's lifestyles, the aging trend is becoming more and more obvious, and the incidence of non-communicable diseases / chronic diseases (such as malignant tumors, cardiovascular and cerebrovascular diseases, diabetes, hypertension, etc.) is constantly increasing. Malignant tumor has become an important health problem in our country, and it is the number one killer of men in our country, and it is second only to heart disease and cerebrovascular disease among women, ranking third 1 . Although the diagnosis and treatment of malignant tumors have made great progress in recent years, for a long time, clinicians often take the same drug at the same dosage for the same malignant tumor according t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 郜恒骏盛海辉
Owner SHANG OUTDO BIOTECH CO LTD
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