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Primer combination used for PARP inhibitor drug susceptibility diagnosing and detection kit

A detection kit and drug-sensitive technology, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of small number of patients, limited application range, non-detection of mutations, etc., and reduce the difficulty , increase versatility, increase the effect of specificity

Inactive Publication Date: 2017-02-22
佳学基因医学技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the above reasons, the current detection kits used for PARP inhibitor drug sensitivity diagnosis often only detect whether the patient's BRCA is mutated, and do not detect mutations in other gene loci, while PARP inhibitors can treat multiple gene loci Tumors caused by mutations, such as TP53, PTEN, and RB1, etc., cause the number of patients screened by existing detection kits to be small, the detection rate of the effectiveness of PARP inhibitors is low, and the scope of use is limited.

Method used

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  • Primer combination used for PARP inhibitor drug susceptibility diagnosing and detection kit
  • Primer combination used for PARP inhibitor drug susceptibility diagnosing and detection kit
  • Primer combination used for PARP inhibitor drug susceptibility diagnosing and detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0046] A primer combination for PARP inhibitor drug sensitivity diagnosis, comprising the following primer sets: M1775K primer set, P151T primer set, G129E primer set, C712R primer set and Q569K primer set, the M1775K primer set includes M1775K-wild-F Primers, M1775K-wild-R primers, M1775K-mutant-F primers and M1775K-mutant-R primers; the P151T primer set includes P151T-wild-F primers, P151T-wild-R primers, P151T-mutant-F primers and P151T-mutant-R primer; The G129E primer set includes G129E-wild-F primer, G129E-wild-R primer, G129E-mutant-F primer and G129E-mutant-R primer; The C712R primer set includes C712R-wild -F primers, C712R-wild-R primers, C712R-mutant-F primers and C712R-mutant-R primers; the Q569K primer set includes Q569K-wild-F primers, Q569K-wild-R primers, Q569K-mutant-F Primers and Q569K-mutant-R primer; the sequence of the M1775K-wild-F primer is shown in SEQ ID NO: 1, the sequence of the M1775K-wild-R primer is shown in SEQ ID NO: 2, the M1775K- The sequence...

Embodiment 2

[0048] A detection kit containing the primer combination described in Example 1, the specific components are as shown in the table below:

[0049] Table 1: Component list of the detection kit

[0050]

[0051]

Embodiment 3

[0053] This embodiment is the use method of the detection kit in embodiment 2 and the result judgment of the example.

[0054] A method for simultaneously detecting BRCA1, BRCA2, TP53, PTEN and RB1 gene mutations using the detection kit, comprising the following steps:

[0055] 1. Take tumor tissue to extract genomic DNA, dissolve the extracted DNA sample with ultrapure water, and adjust the final concentration to 500ng / ul

[0056] 2. According to the sample size to be tested, prepare the reaction mixture according to the following formula, where N is the number of samples

[0057]

[0058] According to the above sequence, add each component into the sample tank in proportion on ice, and mix well to make the test working solution

[0059] 3. In the 96-well plate of fluorescent quantitative PCR, add 48ul test working solution to the A-G wells of each column, and add 1ul 6A, 6B, 6C, 6D, 6E, 6F, 6G solutions to the A-G wells respectively;

[0060] 4. Add 1ul of the positive ...

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Abstract

The invention provides a primer combination used for PARP inhibitor drug susceptibility diagnosing and a detection kit. The primmer combination and the detection kit containing the primmer combination can not only detect mutations of genes of BRCA1 and BRCA2, but also detect mutations of genes of TP53, PTEN and RB1, thus increasing the range and the number of screened patients, and the primmer combination and the detection kit containing the primmer combination can guide cancer patients to use the PARP inhibitor better.

Description

technical field [0001] The invention belongs to the technical field of medical detection supplies, and in particular relates to a primer combination and a detection kit for PARP inhibitor drug sensitivity diagnosis. Background technique [0002] PARP inhibitors can enhance the efficacy of radiotherapy and chemotherapy with alkylating agents and platinum drugs by inhibiting the DNA damage repair of tumor cells and promoting tumor cell apoptosis. Such as olaparib, rucaparib, niraparib and veliparib are all PARP inhibitors. [0003] In addition to improving the efficacy of chemotherapy drugs, PARP inhibitors are approved by the FDA for the treatment of patients with gynecological tumors caused by BRCA1 and BRCA2 mutations. That is, PARP inhibitors as a single drug are also effective for patients with BRCA mutations: PARP and BRCA are the two main genes responsible for repairing DNA mutations in cells, and they protect the stability of gene information when the breast and ovary...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/106C12Q2600/156
Inventor 黄家学杨萍李春芳宋晓琳黄婷婷梁苹苹刘娜
Owner 佳学基因医学技术(北京)有限公司