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Breast cancer early diagnostic kit

A technology for early diagnosis and breast cancer, applied in the field of molecular biology, can solve the problems of lack of specific early diagnosis molecular markers, limited early diagnosis of breast cancer, low signal sensitivity, etc., to improve the cure rate and survival rate, avoid false Negative and false positive results, the effect of improving specificity and accuracy

Inactive Publication Date: 2017-08-01
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are also mRNA early screening technologies for breast cancer-related tumor genes, such as RT-PCR, real-time PCR and fluorescence in situ hybridization, etc., but the current detection is mainly based on the principle of PCR reaction, which has the disadvantages of easy contamination and high false positive rate. , while the traditional RNA fluorescence in situ hybridization itself has the disadvantage of low signal sensitivity
In addition, the lack of specific molecular markers for early diagnosis is also an important factor limiting the early diagnosis of breast cancer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] There are two types of breast cancer early diagnosis kits described in this example, with labeled probes and without labeled probes.

[0025] Wherein, breast cancer early diagnosis kit A with labeled probe mainly includes:

[0026] 1. Capture probe

[0027] The capture probe is composed of three parts, the 5' end to the 3' end are the P1 sequence that is complementary to the mRNA of the corresponding target gene, the spacer sequence, and the P2 sequence that is complementary to the corresponding amplification probe P3 sequence. The P2 sequences in the capture probes of the target genes of the same category are the same. The spacer is used to space the capture probe P2 sequence from the target mRNA, and by setting a spacer sequence of appropriate length inside the probe, it can reduce steric hindrance, improve the efficiency of the hybridization reaction and the specificity of the hybridization reaction . The spacer arm of the capture probe of the present invention is...

Embodiment 2

[0047] Example 2 Using kit A in Example 1 to detect circulating tumor cells in peripheral blood of breast cancer patients

[0048] The formula of described various solutions is as follows:

[0049]

[0050]

[0051] The probe mixture, amplification mixture, and chromogenic mixture in this example all use all the probes in the corresponding gene list of the Breast Cancer Early Diagnosis Kit A with labeled probes in Example 1.

[0052] 1. Sample pretreatment, filtering breast cancer CTCs onto the filter membrane

[0053] 1. Preserve the blood sample in the sample preservation tube with preservation solution, centrifuge at 600×g for 5 minutes, discard the supernatant, and remove the red blood cells.

[0054] 2. Add 4mL PBS and 1mL fixative, vortex to mix, and let stand at room temperature for 8min.

[0055] 3. Sample filtration: transfer the liquid in the sample storage tube to the filter, turn on the vacuum filtration pump to drain the liquid; add 4mL PBS to the storage ...

Embodiment 3

[0105] The selection of embodiment 3 target detection gene quantity and kind

[0106] 1. Design of kit preparation (selection of the number and types of target detection genes)

[0107] The breast cancer screening gene of the kit of the present invention is selected from: MUC1, HER2, hMAM, CEA, COX2, and the corresponding selection is made according to the experimental group. The breast cancer CTC marker gene uses: CK19, SERPINE1, EpCAM and hTERT; the exclusion gene uses: CD45 , CD34, CD105, CD144, CD146 and VWF.

[0108] For the selection of breast cancer screening genes, see Experimental Group 1-4, select one, two, three and five target genes respectively, and compare their detection effects, while breast cancer CTC marker genes and exclusion genes use all target genes , the specific design is shown in Table 7.

[0109] The compositions and quantities of the capture probes, amplification probes and labeling probes, detection methods, etc. of each group of corresponding tar...

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Abstract

The invention relates to a breast cancer early diagnostic kit, which includes a capture prove, an amplification probe and a labeled probe aiming at mRNA detection of target genes. The target genes are a breast cancer screening gene and breast cancer CTC marker gene mRNA. Three molecular markers including a breast cancer screening gene, a breast cancer CTC marker gene and an exclusion gene are used together for early screening of breast cancer, so that the problem of a false-negative result due to expression difference of different individuals and different kinds of breast-cancer-related genes is solved and a false-positive result due to expression enhancement of some related genes in some other non-cancer diseases is excluded. The specificity and accuracy of the detection result are further improved, and misdiagnosis is prevented.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a breast cancer early diagnosis kit. Background technique [0002] Breast cancer is a malignant tumor that occurs in the epithelial tissue of the breast. It is one of the most common malignant tumors in women. The incidence rate accounts for 7% to 10% of all kinds of malignant tumors in the whole body. The incidence of breast cancer is higher in women between the ages of 40 and 60 and before and after menopause, and the incidence is increasing year by year, and it shows a trend of rejuvenation. At present, breast cancer has become a common tumor that threatens women's physical and mental health. The mammary gland is not an important organ to maintain human life activities. Breast cancer in situ is not fatal, but because breast cancer cells lose the characteristics of normal cells, the adhesion between cells changes and they are easy to fall...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 刘苏燕吴诗扬
Owner SUREXAM BIO TECH
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