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Solid phase polymerase chain reaction (PCR) kit for chemotherapeutic medication guide gene detection

A technology for medication guidance and gene detection, applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve the problem of high infection risk, achieve large detection throughput, avoid non-specific amplification, and improve detection sensitivity and specificity. sexual effect

Inactive Publication Date: 2018-02-06
安徽安龙基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Lobaplatin is the third-generation platinum-based antineoplastic drug, mainly used for the treatment of breast cancer, small cell lung cancer, and chronic myelogenous leukemia. The adverse reactions include bone marrow suppression, and thrombocytopenia is the most intense, which often occurs 2 weeks after treatment. After 1 week, it returned to 100,000 / mm3, and the blood picture changes were reversible, but secondary side effects such as thrombocytopenia caused bleeding, leukopenia caused a high risk of infection

Method used

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  • Solid phase polymerase chain reaction (PCR) kit for chemotherapeutic medication guide gene detection
  • Solid phase polymerase chain reaction (PCR) kit for chemotherapeutic medication guide gene detection
  • Solid phase polymerase chain reaction (PCR) kit for chemotherapeutic medication guide gene detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Composition of solid-phase PCR detection kit for guidance of 10 kinds of chemotherapeutic drugs

[0024]The kit consists of a PCR reaction mixture and 12 strip tubes or a 96-well PCR reaction plate, in which the PCR reaction mixture is 5×PCR Buffer (Tris-HCl pH8.5 100mM, KCl 500nM, MgCl2 15nM), dNTPs ( 10mMeach), Mg2+, 12 pairs of primers and hot start DNA polymerase.

[0025] The kit is to seal a layer of paraffin wax mixture on the surface of the above-mentioned PCR reaction mixture, and the paraffin wax mixture is formed by mixing solid paraffin and liquid paraffin according to 1:8 (W / V).

[0026] The reaction system (25ul) of the kit is: PCR reaction solution 23ul, template 2ul.

Embodiment 2

[0028] Kit operation and result determination

[0029] 1. Genomic DNA extraction

[0030] Using paraffin-embedded tissue DNA extraction kit (Tiangen Biochemical Technology, product number: DP331) to extract cancer tissue DNA according to the kit instructions.

[0031] 2. Preparation of reaction system

[0032] Take Chemotherapy Medication Guidance Solid-Phase PCR Reaction Kit 12-tube strip or 96-well PCR reaction plate, and add 2ul of extracted DNA.

[0033] 3. PCR amplification

[0034] Put the PCR tube into an ordinary PCR machine and perform PCR amplification according to the instructions recommended.

[0035] 4. Analysis of PCR products by 2.0% agarose gel electrophoresis

[0036] 5. Interpretation of results

[0037] The result interpretation standard of this kit is as follows Table 2

[0038]

Embodiment 3

[0040] Interpretation of sequencing results and medication guidance

[0041] Using the DNA extracted from the cancer tissue of Example 1 as a template, after amplification with the above-mentioned kit, the sequencing results are obtained, and the corresponding medication instructions are given as follows according to the sequencing results:

[0042]

[0043] The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention should be included in the protection of the present invention. within range.

[0044] sequence listing

[0045] Anhui Anlong Gene Medical Laboratory Co., Ltd.

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Abstract

The invention discloses a solid phase polymerase chain reaction (PCR) kit for chemotherapeutic medication guide gene detection, and a detection method, belonging to the technical field of PCR application. The invention also relates to a primer group for detecting ten chemotherapeutic medication guide; the primer group consists of GSTP1 SNP-I105V A313G, MTHFR SNP-C677T, MDR1 G2677AT, XRCC1 SNP-R399Q, CDA G208A, MDR1 SNP-C3435T, CDA A79C, CYP1B1C SNP-4326C / G, ERCC1 SNP-C118T, DPYD IVS14_1G / A; the solid phase PCR amplification kit can be used for detecting the ten gene sites at a time; computer amplification can be realized only by adding a corresponding nucleic acid template into the kit again; the PCR product, obtained after amplification is completed, is subjected to Sanger sequencing; chemotherapeutic medication guide can be realized according to the result of the Sanger sequencing. Compared with the prior art, the solid phase PCR kit adopts a specific primer sequence, so that the reliability of the detection results is ensured; the detection method is simple to operate, time-saving and labor-saving; the solid phase PCR kit is high in detection throughput and low in reagent consumable cost, can be directly used for detecting extracted nucleic acid, is low in requirements for a detection platform and detection personnel, and can be widely popularized in the clinical front line.

Description

technical field [0001] The invention relates to the application field of gene detection, in particular to a solid-phase PCR kit for chemotherapy drug guidance gene detection. Background technique [0002] Tumor is a disease caused by gene mutation. Genetic testing can find out the gene mutation in the tumor. Combined with conventional pathological diagnosis, it can more accurately analyze the type, grade, molecular type, drug sensitivity and other information of the tumor, helping clinicians to have a more accurate diagnosis. Tailored treatment plan. Therefore, please listen carefully to the advice of clinicians, carry out relevant genetic testing under the guidance of doctors, and receive corresponding treatment. [0003] With the development of individualized treatment, more and more clinicians and patients realize that the efficacy and side effects of drugs can be predicted by genetic testing before the treatment of tumors, so as to choose the tumor that suits them, has ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/106C12Q2600/156C12Q2600/16
Inventor 韦玉军李航高雅丽苏军吴远航
Owner 安徽安龙基因科技有限公司
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