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Kit for platinum drug medication guidance

A platinum drug and kit technology, which is applied in the field of kits for guiding the administration of platinum drugs, can solve problems such as differences in repair ability, limit the scope of use of cisplatin, and affect the sensitivity of platinum drugs, and achieve simple, safe and reasonable operation. Effective individualized drug delivery, improved targeting and predictable outcomes

Inactive Publication Date: 2015-08-26
北京奥维森基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The second-generation platinum-based anticancer drug carboplatin is very similar in structure to cisplatin, and has similarities in its mechanism of action and drug resistance. Both are used to treat ovarian cancer, non-small cell lung cancer, head and neck cancer, and cervical cancer. , lymphoma, etc., but the use of cisplatin and carboplatin is limited due to toxicity and drug resistance
DNA repair ability is related to the change of XRCC1 gene phenotype. Due to the existence of single nucleotide polymorphism (SNP), the activity of XRCC1 in different individuals is different, which may lead to differences in repair ability among different individuals, thus affecting the response to platinum drugs. sensitivity

Method used

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  • Kit for platinum drug medication guidance
  • Kit for platinum drug medication guidance
  • Kit for platinum drug medication guidance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1, the preparation of specific primer pair, positive control and negative control

[0027] 1. Preparation of specific primer pairs

[0028] The specific primer pair for detecting the C118T point mutation of ERCC1 gene consists of F1 and R1.

[0029] F1 (sequence 1 of the sequence listing): 5'-TGGCGAATATGGTGACAC-3';

[0030] R1 (SEQ ID NO: 2 of the Sequence Listing): 5'-CTCATAGAACAGTCCAGAACA-3'.

[0031] The specific primer pair for detecting the R194W point mutation of the XRCC1 gene consisted of F2 and R2.

[0032] F2 (sequence 3 of the sequence listing): 5'-GTGAAGGAGGAGGATGAGA-3';

[0033] R2 (SEQ ID NO: 4 of the Sequence Listing): 5'-CTGGCTGTGACTATGAAGG-3'.

[0034] The specific primer pair for detecting the I105V point mutation of GSTP1 gene consisted of F3 and R3.

[0035] F3 (sequence 5 of the sequence listing): 5'-CATCCTTCCACGCACATC-3';

[0036] R3 (SEQ ID NO: 6 of the Sequence Listing): 5'-CGTTACTTGGCTGGTTGAT-3'.

[0037] The specific primer pa...

Embodiment 2

[0045] Embodiment 2, the sensitivity detection of specific primer pair

[0046] 1. Sensitivity detection of primers to formazan

[0047] Using the positive control plasmid A as a template, the primer pair A was used for PCR amplification.

[0048] The total volume of the PCR amplification system is 25 μl. The PCR amplification system contains 0.5μM F1, 0.5μM R1 and 1U hot-start DNA polymerase. The PCR amplification systems contained 10ng / μl, 5ng / μl, 2.5ng / μl, 1ng / μl, 0.5ng / μl, 0.25ng / μl, 0.1ng / μl or 0.05ng / μl positive control plasmid A respectively.

[0049] The PCR amplification program is: 95°C for 5 minutes; 30 cycles of 95°C for 30 seconds, 53°C for 30 seconds, and 72°C for 30 seconds; 72°C for 10 minutes; 15°C, hold.

[0050] The PCR amplification products were subjected to 2% agarose gel electrophoresis, and the results were shown in figure 1 . figure 1 Among them, t1 to t8 successively represent the concentration of positive control plasmid A is 10ng / μl, 5ng / μl, 2....

Embodiment 3

[0064] Embodiment 3, the application of specific primer pair

[0065] 1. Collect blood samples from 8 volunteers who gave informed consent, and extract genomic DNA.

[0066] 2. Use each genomic DNA extracted in step 1 as a template, and use primers to perform PCR amplification on A (in the initial PCR amplification system, the content of genomic DNA is 30ng); set a positive control with positive control plasmid A instead of genomic DNA Treatment, set the negative control treatment with negative control instead of genomic DNA.

[0067] The total volume of the PCR amplification system is 25 μl, containing 0.5 μM F1, 0.5 μM R1 and 1 U hot-start DNA polymerase.

[0068] The PCR amplification program is: 95°C for 5 minutes; 30 cycles of 95°C for 30 seconds, 53°C for 30 seconds, and 72°C for 30 seconds; 72°C for 10 minutes; 15°C, hold.

[0069] 3. Use each genomic DNA extracted in step 1 as a template, and use primer pair B to carry out PCR amplification (in the initial PCR amplif...

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Abstract

The invention discloses a kit for platinum drug medication guidance. The invention protects a primer group. The primer group comprises a primer pair A, a primer pair B and a primer pair C. The primer pair A comprises a single chain DNA molecule shown in the sequence 1 and a single chain DNA molecule shown in the sequence 2. The primer pair B comprises a single chain DNA molecule shown in the sequence 3 and a single chain DNA molecule shown in the sequence 4. The primer pair C comprises a single chain DNA molecule shown in the sequence 5 and a single chain DNA molecule shown in the sequence 6. The invention also protects a kit for identification of C118T point mutation of an ERCC1 gene, R194W point mutation of a XRCC1 gene and I105V point mutation of a GSTP1 gene. The kit comprises the primer group. The kit can be used for detecting three platinum drug sensitivity-related SNP sites, can be further used for platinum drug individual use guidance and improves clinical treatment specificity and predictability.

Description

technical field [0001] The invention relates to a kit for guiding the administration of platinum drugs. Background technique [0002] At present, the common problems in clinical chemotherapeutics are tumor drug resistance and side effects of chemotherapeutic drugs. The future development direction of chemotherapy is to realize the individualized treatment mode according to the response of the tumor and the side effects of the host. Platinum anticancer drugs (mainly refer to cisplatin, carboplatin and oxaliplatin; the full name of cisplatin is cis-dichlorodiammine platinum, the full name of carboplatin is cis-diaminocyclobutane platinum, oxaliplatin The full name of platinum is L-trans diaminocyclohexane oxalate platinum), which is an anticancer drug with high activity against various cancers in clinical application. Cisplatin (cisplatin) has been widely used in cancer chemotherapy since it was discovered in the 1970s that it can inhibit the growth of tumor cells. Cisplati...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/106C12Q2600/156
Inventor 文洁
Owner 北京奥维森基因科技有限公司
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