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Ovarian cancer individualized treatment detection reagent case and application thereof

A detection kit and technology for ovarian cancer, applied in the determination/testing of microorganisms, biochemical equipment and methods, fluorescence/phosphorescence, etc., can solve the problems of chemotherapy drugs, drug resistance, treatment failure, etc.

Inactive Publication Date: 2015-05-06
DAAN GENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current clinical application of platinum-based combined chemotherapy has a good therapeutic effect on ovarian cancer, but most patients with advanced ovarian cancer will eventually relapse and develop chemotherapeutic drug resistance, resulting in treatment failure

Method used

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  • Ovarian cancer individualized treatment detection reagent case and application thereof
  • Ovarian cancer individualized treatment detection reagent case and application thereof
  • Ovarian cancer individualized treatment detection reagent case and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1: Preparation method of BRCA1 gene probe for molecular marker detection of ovarian cancer

[0092] (1) Cloning screening: select clones containing the target gene BRCA1 and the sequences at both ends of RP11-948G15 and RP11-831F13.

[0093] (2) Clone culture and identification: Purchase the corresponding clone Invitrogen RPCI11.C, add 50ul to 500ml of TB culture medium (chloramphenicol resistant), shake culture in a shaker at 37°C for 24 to 48 hours; use the bacterial solution STS primer pair for clone identification. RP11-948G15: STS primer pair: upstream primer 5'-CTTGCACCTATAATCCCAG-3' and downstream primer 5'-AGTCTTCTTGTCTTGTGGC-3', PCR amplification conditions: 95°C for 5 minutes; (94°C for 30 seconds, 55°C for 30 seconds, 72°C for 45 seconds) × 35 cycles; 72°C for 7 minutes. The amplified product was verified by electrophoresis, and the result showed a bright band around 128 bps. RP11-831F13STS primer pair: upstream primer 5'-AAACATGTTCCTCCTAAGGTGCTTT-3' an...

Embodiment 2

[0104] Example 2: Preparation method of ovarian cancer molecular marker detection agent PTEN gene probe

[0105] (1) Cloning screening: Select clones containing the target gene PTEN and the sequences at both ends of RP11-113H4, CTD-2557P6 and RP11-765C10.

[0106] (2) Clone culture and identification: RP11-113H4: STS primer pair: upstream primer 5'-AGAAAGACAGACAAAAAGATGAGG-3' and downstream primer 5'-GAGTTGCCAGTGTGCTTTAC-3', PCR amplification conditions: 95°C for 5 minutes; (94°C) 30 seconds, 58°C for 30 seconds, 72°C for 45 seconds) × 35 cycles; 72°C for 7 minutes. The amplified product was verified by electrophoresis, and the result showed a bright band around 188bps. CTD-2557P6 STS primer pair: upstream primer 5'-GTTTTTAGTGAGGGCTGGTGAAA-3' and downstream primer 5'-ATTTCCCTTTGGAAAGTGCTGTT-3', PCR amplification conditions are: 95°C for 5 minutes; (94°C for 30 seconds, 58°C for 30 seconds, 72 ℃ 45 seconds) × 35 cycles; 72 ℃ 7 minutes. The amplified product was verified by electr...

Embodiment 3

[0117] Example 3: Preparation method of WT1 gene probe for ovarian cancer molecular marker detection agent

[0118] (1) Cloning screening: select clones containing the target gene WT1 and the sequences at both ends of RP11-1037K14, CTD-2643K15 and RP11-195J14.

[0119] (2) Clone culture and identification: RP11-1037K14: STS primer pair: upstream primer 5'-AGCTCAAGTGGACAGATGTACAGG-3' and downstream primer 5'-TGCAATTAGCACGACCATGATAC-3', PCR amplification conditions: 95°C for 5 minutes; (94°C) 30 seconds, 56°C for 30 seconds, 72°C for 45 seconds) × 35 cycles; 72°C for 7 minutes. The amplified product was verified by electrophoresis, and the result showed a bright band around 323bps. CTD-2643K15STS primer pair: upstream primer 5'-GTTGTCCTTTTCAGCATTGC-3' and downstream primer 5'-GCAGGGTTTCATCCTCGG-3', PCR amplification conditions are: 95℃ for 5 minutes; (94℃ for 30 seconds, 58℃ for 30 seconds, 72℃ 45 seconds) × 35 cycles; 72°C for 7 minutes. The amplified product was verified by elec...

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Abstract

The invention relates to an ovarian cancer individualized treatment detection reagent case and application of the ovarian cancer individualized treatment detection reagent case, in particular to the ovarian cancer individualized treatment. The ovarian cancer individualized treatment detection reagent case and the application of the ovarian cancer individualized treatment detection reagent case comprise development and progression, staging and grading, forecast treatment and preparation of related molecular marker detection probes. The related molecular marker detection probes can be used for guiding the planning of an ovarian cancer individualized treatment schedule, evaluating the treatment effect, monitoring recurrence and metastasis, estimating prognosis, and the like.

Description

Technical field [0001] The invention relates to a detection agent for individualized treatment of ovarian cancer and its application in the detection of ovarian cancer. Specifically, it relates to the individualized treatment of ovarian cancer, including the preparation of related molecular marker detection probes such as occurrence and development, staging and grading, and predicting therapeutic effect. The ovarian cancer molecular marker detection probe involved in the present invention can be used to guide the formulation of an individualized treatment plan for ovarian cancer, evaluate the treatment effect, monitor recurrence and metastasis, evaluate prognosis, and the like. Background technique [0002] Ovarian cancer is a malignant tumor that occurs in ovarian tissue. Its incidence is second only to cervical cancer and uterine body cancer, and it accounts for the third place among female malignant tumors. Because ovarian cancer is located in the pelvic cavity, there are few...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 李明何瑰陈华云
Owner DAAN GENE CO LTD
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