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A breast cancer screening kit

A technology of breast cancer and kits, which is applied in the direction of instruments, biochemical equipment and methods, and analytical materials, and can solve the problems of unsatisfactory screening results

Active Publication Date: 2019-11-05
成竞梁
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, breast cancer screening methods commonly used at home and abroad include mammography (mammography), ultrasound imaging, clinical breast examination and magnetic resonance imaging, etc., but the screening results are not satisfactory. Looking for new and reliable methods Screening methods are of great significance for the detection of early breast cancer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 RAPD method specifically amplifies the markers for detecting breast cancer of the present invention (nucleotide sequences shown in SEQ ID NO.1, 2)

[0035] RAPD (Random Amplified Polymorphic DNA)—that is, random amplified polymorphic DNA, is mainly used for molecular detection when the genome information of animals and plants is unknown. RAPD technology relies on PCR, and selects 10 base single-stranded random primers to perform PCR amplification on the DNA of organisms, populations or species genomes to detect differences in genetic loci.

[0036] 1. Experimental method

[0037] (1) Tumor tissue DNA acquisition

[0038] 1. Take 5 pairs of breast cancer tissue and adjacent normal tissue (tissue area 2 cm next to the lesion) (Table 1). Take 5 mg of breast cancer tissue, cut it into pieces, put it in a glass homogenizer, add 2-3ml of nuclei lysis buffer, and homogenize until no tissue lumps are seen;

[0039]2. Transfer all the homogenizer into a 5ml centrifu...

Embodiment 2

[0153] Example 2 Relationship between expression levels of nucleotide sequences shown in SEQ ID NO.1, 2 and breast cancer (genome level)

[0154] 1. Experimental method

[0155] (1) After sequencing analysis, according to the determined sequence, use Primer3.0 software (http: / / frodo.wi.mit.edu / primer3) to design specific primers and synthesize primers (Table 3). These primers are used to amplify Genomic DNA of tumors and normal people, GAPDH as internal control, they can be used for semi-quantitative PCR.

[0156]

[0157] BC10-1L (SEQ ID NO.3), BC10-1R (SEQ ID NO.4);

[0158] BC31-2L (SEQ ID NO.5), BC31-2R ​​(SEQ ID NO.6);

[0159] GAPDH5G (SEQ ID NO. 15), GAPDH3G (SEQ ID NO. 16).

[0160] At the same time, real-time fluorescent quantitative PCR analysis primers were designed (Table 4). The primers in Table 4 are used to amplify the genomic DNA of tumors and normal people, β-actin is used as an internal reference, and the Taqman probe number (Universal Probe Library, R...

Embodiment 3

[0194] Example 3 Relationship between expression levels of DPEP1 and PHKG2 and breast cancer (mRNA level)

[0195] 1. Experimental method

[0196] (1) Extraction of total RNA from cells and tumor tissues

[0197] Total cellular RNA extraction

[0198] 1. Use miRNeasy Mini Kit or RNeasy Mini Kit purchased from QIAGEN (Cat#217004 or Cat No.74104)

[0199] 2. Take well-grown breast cancer cells with a density of more than 95% (6-well plate), absorb the residual liquid of the medium, and rinse twice with pre-cooled 1×PBS;

[0200] 3. Add 700 μl of QIAzol Lysis (miRNeasy Mini Kit), draw 5 times with a 1ml syringe, and let stand for 5 minutes;

[0201] 4. Add 140 μl of chloroform, shake vigorously for 15 seconds and then let stand at room temperature for 2-3 minutes;

[0202] Centrifuge at 10,000 rpm at 5.4°C for 15 minutes;

[0203] 6. Carefully draw the supernatant into another EP tube, add 1.5 times the volume of absolute ethanol, and turn it upside down several times;

[0...

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Abstract

The invention provides a nucleotide sequence as shown in SEQ ID No. 1 or SEQ ID No. 2, application of the nucleotide sequence and a breast cancer screening kit. The nucleotide sequence and the breast cancer screening kit can be used to assist clinical breast cancer diagnosis and are promising in clinical application prospect.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a breast cancer screening kit. Background technique [0002] Breast cancer (breast cancer, BC) is the most common malignant tumor in women, accounting for 7-10% of all kinds of malignant tumors in the whole body. Eating habits, environmental carcinogens, etc. are all key factors that induce breast cancer. [0003] At present, there is still no effective treatment for breast cancer. Breast cancer screening is recognized as the main method that can effectively improve the survival rate of breast cancer in women. Breast cancer has been listed by WHO as one of the cancer types that should be screened in the population. Reasonable screening can detect breast cancer early, improve the cure rate, increase the chance of "breast-conserving" surgery, reduce postoperative adjuvant therapy, save medical expenses, and improve the quality of life of patients. [0004] At prese...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12Q1/6886G01N33/574
CPCC12Q1/6886C12Q2600/158G01N33/57415G01N33/57484
Inventor 付尚颐付俊江成竞梁魏春莉杨璐全
Owner 成竞梁
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