SNP marker associated with curative effect of novel adjuvant chemotherapy of breast cancer and application of SNP marker
A breast cancer and marker technology, applied in the field of SNP markers, can solve the problems of low pCR rate, NACT insensitivity, delayed local treatment, etc., to improve sensitivity and specificity, facilitate detection, improve pathological complete remission rate and Effects on long-term survival
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Embodiment 1
[0048] This embodiment is the collection of samples and the arrangement of sample data.
[0049] From September 2015 to November 2016, the inventor collected a large number of blood samples from breast cancer chemotherapy patients at Huashan Hospital Affiliated to Fudan University. After sorting out the sample data, the inventor selected 90 samples that met the following criteria: (1) Females aged 18-70 years; (2) Breast cancer patients confirmed by pathological biopsy or cytology; (3) Measurable tumor lesions confirmed by mammography or B-ultrasound; (4) Accepted 4 cycles of neoadjuvant chemotherapy with cyclophosphamide (C) + epirubicin (E) + paclitaxel or docetaxel (T); (5) left / right mastectomy after neoadjuvant chemotherapy; (6) ) General physical condition score PS 0-2; (7) No major organ dysfunction, blood routine, liver and kidney function and heart function are basically normal; (8) Complete follow-up information can be obtained; Demographic and clinical data, etc. ...
Embodiment 2
[0051] This embodiment is the peripheral blood DNA extraction, genotype detection and result analysis.
[0052] Among the 90 eligible breast cancer patients in the above-mentioned embodiment, their disease state and physiological state were balanced and comparable, and there were statistical differences.
[0053] The specific steps are:
[0054] 1) Extract 200 μl fresh blood, add 20ul Proteinase K solution, and mix well;
[0055] 2) Add 200 μl buffer GB, mix thoroughly by inversion, place at 70°C for 10 minutes, and briefly centrifuge to remove water droplets on the inner wall of the tube cap;
[0056] 3) Add 200 μl of absolute ethanol, shake and mix well for 15 sec, and briefly centrifuge to remove water droplets on the inner wall of the tube cap;
[0057] 4) Add the solution and flocculent precipitate obtained in the previous step into an adsorption column CB3 (the adsorption column is placed in a collection tube), centrifuge at 12000rpm (~13400×g) for 30sec, pour off the ...
Embodiment 3
[0067] This example is the production of a kit for detecting or predicting the curative effect of paclitaxel-based neoadjuvant chemotherapy for breast cancer.
[0068] The production and operation process of the SNP kit is based on the Sequenom Mass ARRAY genotyping platform detection technology. The kit contains a set of specific amplification primers for SNP markers (comprising the following primers: the rs2981578 amplification primer whose sequence is SEQ ID No:1 and SEQ ID No:2 and / or whose sequence is SEQ ID No:3 and SEQ ID No:4 rs1966265, see Table 1 for details), and common reagents required by corresponding PCR techniques, such as: dNTPs, MgCl 2 , double distilled water, etc., these commonly used reagents are well known to those skilled in the art, and there may also be standards and controls (such as standards for genotype determination and blank controls, etc.). The value of this kit is that it only needs peripheral blood and no other tissue samples, detects the SNP...
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