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Kit for evaluating treatment sensitivity and/or drug resistance of apatinib

An apatinib and sensitivity technology, applied in the field of kits for evaluating the sensitivity and/or drug resistance of apatinib treatment, to achieve the effects of short detection time, simple operation and low price

Active Publication Date: 2022-04-26
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Apatinib inhibits tumor neovascularization and is approved for the third-line and above treatment of advanced gastric cancer or gastroesophageal junction adenocarcinoma. The drug also shows good anti-tumor activity in various other tumors. The application in cancer is still in the preliminary exploration stage

Method used

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  • Kit for evaluating treatment sensitivity and/or drug resistance of apatinib
  • Kit for evaluating treatment sensitivity and/or drug resistance of apatinib
  • Kit for evaluating treatment sensitivity and/or drug resistance of apatinib

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] A kind of method of the PEX3 gene of embodiment 1 detection

[0055] 1. Primer design

[0056] According to the Peroxisomal Biogenesis Factor 3 (PEX3, NCBI: NM_003630.3) gene and internal reference gene (GAPDH, NCBI: NM_001256799.3) sequence registered in GenBank, design specific primers and probes, and use the BLAST function of NCBI to compare Specificity was initially identified, and the sequences of primers and probes are shown in Table 1.

[0057] Table 1:

[0058]

[0059] 2. Real-time fluorescent quantitative PCR amplification and detection of PEX3 gene

[0060] (1) Reagent preparation: 18 μl of PCR reaction solution, 2 μl of enzyme mixture, mix well and set aside.

[0061] Wherein the PCR reaction solution is composed of primers (SEQ ID NO.1~2 and SEQ ID NO.4~5), probes (SEQ ID NO.3 and SEQ ID NO.6) and PCR buffer in Table 1, wherein the primers The final concentration of the amplification system is 0.2 μmol / L, the final concentration of the probe amplific...

Embodiment 2

[0074] A kind of test kit of the PEX3 gene of embodiment 2 detection

[0075] 1. Composition

[0076] 1 tube of PEX3 PCR reaction solution, consisting of primers, probes and PCR buffer, wherein the final concentration of primer amplification system is 0.2 μmol / L, the final concentration of probe amplification system is 0.1 μmol / L, and the final concentration of PCR buffer is 1×, the specific primers and probes are as follows:

[0077] PEX3 gene upstream primer: 5'-GACCTGCAACATGGTAACTCT-3' (SEQ ID NO.1)

[0078] PEX3 gene downstream primer: 5'-CACTTGCTCCATTGTCAACA-3' (SEQ ID NO.2)

[0079] PEX3 gene probe: 5'-FAM-ACTGCAAACTGAATGGATCTGTCCG-BHQ1-3'(SEQ ID NO.3)

[0080] Internal reference gene upstream primer: 5'-AAAACCTGCCAAATATGATGACA-3' (SEQ ID NO.4),

[0081] Internal reference gene downstream primer: 5'-TGAAGTCAGAGGAGACCACC-3'(SEQ ID NO.5),

[0082] Internal reference gene probe: 5'-VIC-CAGTGTAGCCCAGGATGCCCTT-BHQ1-3'(SEQ ID NO.6);

[0083] 1 tube of PEX3 enzyme mixture...

Embodiment 3

[0096] The kit of the PEX3 gene detected in embodiment 3 detects the amplification efficiency of PEX3 gene and internal reference gene

[0097] 1. Experimental method

[0098] Select the RNA of colorectal cancer tissue samples with higher concentrations of PEX3 gene and internal reference gene, respectively, and prepare 10-fold gradient dilutions into 5 gradients to prepare quantitative standards as templates, and use the kit in Example 2 for detection. Positive controls were used for quality control.

[0099] 2. Experimental results

[0100] The amplification curves of the FAM and VIC channels of the negative quality control are all obvious S-shaped curves, and the Ct (objective)-Ct (internal reference) of the negative quality control is ≤ 3; the amplification of the FAM and VIC channels of the positive quality control The curves are all obvious S-shaped curves, and the Ct (objective)-Ct (internal reference) of the positive quality control product is >3. Both negative and ...

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Abstract

The invention discloses a kit for evaluating treatment sensitivity and / or drug resistance of apatinib, and provides a kit which can help a clinician to determine whether a colorectal cancer patient is suitable for fluorouracil drug treatment or not according to the detection condition of PEX3 gene mRNA in tumor tissue of the colorectal cancer patient. The kit provides a basis for formulating a personalized treatment scheme, is simple and convenient to operate, short in detection time, capable of realizing high-throughput detection and low in price, is mainly applied to detection of the PEX3 gene, and can be used for guiding clinicians to take apatinib for colorectal cancer patients.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a kit for evaluating the treatment sensitivity and / or drug resistance of apatinib. Background technique [0002] Colorectal cancer (CRC) is one of the most common malignant tumors of the digestive tract with the third highest incidence in the world, and its morbidity and mortality are increasing year by year. According to the latest research data on cancer in China, about 376,000 people are diagnosed with CRC every year, and its incidence rate ranks fifth among all malignant tumors; about 191,000 people die of CRC every year, and its case fatality rate ranks 5th among all malignant tumors. It is also ranked 5th. [0003] At present, there are various kinds of chemical drugs for the treatment of colorectal cancer, and some studies have found that apatinib has a better clinical effect on advanced cancer. Apatinib can highly selectively compete for the ATP-binding site of intr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/6886C12Q1/686C12Q2600/106C12Q2600/158C12Q2600/166C12Q2521/107C12Q2545/101C12Q2545/113C12Q2563/107C12Q2527/127Y02A50/30
Inventor 赵丽瑛马强
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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