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Mutant gene group, library and kit for evaluating risk of female malignant tumor

A technology for malignant tumors and mutated genes, applied in the field of biomedical molecular detection, which can solve the problems of short time, many sites, and large number of people who can be detected.

Pending Publication Date: 2020-09-08
AIR FORCE MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

FISH detection method can directly and accurately determine whether the target gene is amplified, but the detection process is cumbersome and expensive; gene chip technology can detect a large number of people, many sites, and a short time, but it is only used for published gene sites. High price; Real-time PCR method is sensitive, but has high requirements on the experimental system, and the repeatability of the results is poor
As more and more susceptibility genes related to female malignant tumors have been identified, none of the existing detection methods can meet the clinical needs of early screening, early diagnosis and early treatment of female malignant tumors

Method used

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  • Mutant gene group, library and kit for evaluating risk of female malignant tumor
  • Mutant gene group, library and kit for evaluating risk of female malignant tumor
  • Mutant gene group, library and kit for evaluating risk of female malignant tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0377] This embodiment discloses the construction method of the DNA library of the present invention, specifically:

[0378] 1. DNA extraction and dilution: The concentration of the extracted DNA was detected by Nanodrop, and the DNA of all samples was diluted to 30 ng / μl, and the final volume of each diluted sample was 12 μl.

[0379] 2. Operate on ice for PCR amplification. The preparation of the experimental system is as follows:

[0380]

[0381] The system was prepared according to the number of samples to be tested. A total of 3 sets of primer pools were designed for 25 genes, and each sample was amplified with 3 tubes of different primer pools in the first round. When dispensing, first divide the above-prepared MIX into 3 tubes, add 11×sample number μl of primers to each tube, mix well, take 16 μl into PCR tubes corresponding to the number of samples, and dilute each sample Take 4 μl of DNA and add it, and mark the primer pool and sample number at the same time.

...

Embodiment 2

[0414] Template preparation (Ion PGM TM Hi-Q TM View OT2 Kit, Thermo Fisher, Cat. No. A29900)

[0415] 1) Turn on the power switch on the back of IonOne Touch2: Run the Clean program once (for 15 minutes), press Open LID on the touch screen, after the centrifuge is turned on, use 2 Ion One TouchTM Breaking Solution, and then 1 Ion One TouchTM Recovery Put the Router into the centrifuge, and then cover the centrifuge cover.

[0416] 2) Remove the used cleaning adapter, put it into the board (the hole is aligned with the left hole of the instrument), and push the wrench to cover the thermal cover. Pass the hose under the wrench and secure it to the pinch valve snap.

[0417]3) Insert the disposable syringe, make sure that the syringe pillow can touch the Router when it is pressed down, and it can spring back after being released. 4) Every time you start: Use a new Ion PGMTM Hi-Q OT2 Kit, discard the IonOneTouchTM Sipper and Ion OneTouchTM Tubes used in the previous kit, put ...

Embodiment 3

[0470] On-machine sequencing, the kit used is Ion PGM TM Hi-Q TM View Sequencing Kit, Thermo Fisher, Cat. No. A30044.

[0471] Preparation

[0472] 1. New Wash2 Bottles processing:

[0473] 1) Transfer the Wash2 bottle to 2L fresh 18MΩ ultrapure water to the scale;

[0474] 2) Add Wash2 Bottle Conditioning Solution, mix up and down 5 times;

[0475] 3) Treat at room temperature for at least 8 hours.

[0476] 2. Use a special pure water bucket to receive fresh 18MΩ ultrapure water.

[0477] 3. Cleaning

[0478] 1) Wash the 3 sequencing vials (Wash1, Wash2 and Wash3) 3 times each.

[0479] 2) Wash 3 wash bottles (Wash1, Wash2 and Wash3) 3 times each.

[0480] 3) Wash 3 chlorine wash bottles (Wash1, Wash2 and Wash3) 3 times each.

[0481] 4) Wash the 1L serum bottle 3 times.

[0482] 3. Store water

[0483] 1) Fill the sequencing bottle Wash2 with 2L of fresh 18MΩ ultrapure water to the mark on the bottle body.

[0484] 2) Washing bottle Wash1 connects 250m1 fresh 18M...

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Abstract

The invention discloses a mutant gene group, library and kit for evaluating female malignant tumor risk, and belongs to the technical field of biomedical molecular detection. The mutant gene group forevaluating the risk of female malignant tumors comprises one or more selected from a group consisting of the following 25 genes: ATM, BRCA1, BRCA2, BRIP1, CDH1, CHEK2, EPCAM, MLH1, MSH2, MSH6, NBN, NF1, PALB2, PMS2, PTEN, RAD51C, RAD51D, STK11, TP53, BARD1, MRE11A, MUTYH, RAD50, XRCC2 and FANCC5. According to the invention, female malignant tumor related genetic markers can be widely screened andinspected, accuracy is higher, and a basis is provided for prediction, prevention and diagnosis of female malignant tumor onset risks.

Description

technical field [0001] The invention belongs to the technical field of biomedical molecular detection, and specifically relates to a mutation gene group, a library and a kit for evaluating the risk of female malignant tumors. Background technique [0002] Breast cancer is a malignant tumor originating from breast epithelial tissue, which accounts for a quarter of female tumors and seriously endangers women's health. In recent years, with the increase of early screening and the improvement of treatment methods, the survival and prognosis of breast cancer have been significantly improved, but the overall situation is still not ideal. According to statistics, in 2012, there were approximately 1.7 million new cases of breast cancer and 520,000 breast cancer-related deaths worldwide. By 2018, the number of new breast cancer cases had increased to 2.1 million and the death toll was approximately 630,000. With changes in lifestyle and reproductive patterns, the incidence and morta...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858C40B40/06C40B50/06
CPCC12Q1/6886C12Q1/6858C40B40/06C40B50/06C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2535/122
Inventor 徐盈杨红张建芳李佳郭芬芬宋婷婷黎昱李春艳郑娇
Owner AIR FORCE MEDICAL UNIV
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