Identification kit for circulating tumor cells of breast cancer

A breast cancer cell and tumor cell technology, applied in breast cancer circulating tumor cell identification kits, medical and biological fields, can solve the problems of contaminated blood samples, false positives, unable to detect CTCs that lose epithelial antigens, etc., to improve the accuracy and reliability, the effect of improving accuracy

Inactive Publication Date: 2017-06-20
SUREXAM BIO TECH
View PDF3 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although CTCs detection has been widely used in breast cancer, the CellSearch detection system approved by the US FDA for clinical application can also be used for the detection and counting of CTCs in the peripheral blood of breast cancer patients, but the CellSearch detection system is based on epithelial cell markers. Target counts CTCs, so CTCs missing epithelial antigens cannot be detected
Like other cancer types, the identification and typing of CTCs in breast cancer patients currently mainly rely on the detection of general CTCs epithelial and mesenchymal marker molecules, because there may be a certain number of non-neoplastic epithelial cells in peripheral blood, and blood sampling may Contamination of blood samples by normal epithelial cells, coupled with differential expression of some epithelial and mesenchymal markers in different cancer types, can lead to some false positive and false negative test results
At present, there are no stable products for the identification and typing of CTCs in breast cancer patients at home and abroad

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Identification kit for circulating tumor cells of breast cancer
  • Identification kit for circulating tumor cells of breast cancer
  • Identification kit for circulating tumor cells of breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] There are two types of breast cancer circulating tumor cell identification kits described in this example, with labeled probes and without labeled probes.

[0043] Wherein, breast cancer circulating tumor cell identification kit A with labeled probe mainly includes:

[0044] 1. Capture probe

[0045] The capture probe consists of three parts, from the 5' end to the 3' sequence is the sequence P1 that is complementary to the mRNA of the corresponding marker gene, the spacer sequence, and the P2 sequence that is complementary to the corresponding amplification probe P3 sequence, the same category The P2 sequences in the capture probes of the marker genes 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 o...

Embodiment 2

[0073] Example 2 Using kit A in Example 1 to detect circulating tumor cells in peripheral blood of breast cancer patients

[0074] The formula of described various solutions is as follows:

[0075]

[0076]

[0077] 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 Circulating Tumor Cell Identification Kit A with Labeled Probes in Example 1.

[0078] 1. Sample pretreatment, filtering breast cancer CTCs onto the filter membrane

[0079] 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.

[0080] 2. Add 4ml of PBS and 1ml of fixative, vortex to mix, and let stand at room temperature for 8min.

[0081] 3. Sample filtration: transfer the liquid in the sample storage tube to the filter, turn on the vacuum filtration pump to pump out the liqu...

Embodiment 3

[0141] Example 3 Using the kit A in Example 1 to detect breast cancer cell lines

[0142] 1. Selection of cell lines

[0143] The breast cancer cell marker gene of the kit of the present invention is selected from: two or more of HER2, SCGB2, MUC1, MGB1; the epithelial cell marker gene is selected from: one or more of EpCAM, CDH1, KRT7, KRT8, KRT18, KRT19 More than one; Mesenchymal cell marker gene selected from one or more of FN1, CDH2, VIMENTIN, FOXC1, TWIST1; Leukocyte marker gene is CD45. The various marker genes in the breast cancer cell marker gene, epithelial cell marker gene, mesenchymal cell marker gene and leukocyte marker gene selected in the present invention are the specific genes on breast cancer CTCs obtained by the inventor through a large number of experiments and statistical analysis. The expressed gene has good specificity and accuracy for the detection of breast cancer cells.

[0144] In this example, the epithelial-mesenchymal mixed breast cancer cell li...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Membrane pore sizeaaaaaaaaaa
Login to view more

Abstract

The invention relates to an identification kit for circulating tumor cells of breast cancer. The identification kit comprises capturing probes, amplification probes and labeling probes aiming at marker gene mRNAs, wherein the marker gene mRNAs are selected from breast cancer cell marker gene mRNAs selected from at least two of HER2, SCGB2, MUC1 and MGB1, epithelial cell marker gene mRNAs selected from at least one of EpCAM, CDH1, KRT7, KRT8, KRT18 and KRT19, and mesenchymal cell marker gene mRNAs selected from at least one of FN1, CDH2, VIMNETIN, FOXC1 and TWIST1. With the identification kit, the false positive result caused by the factors including non-tumor epithelial cells in the blood and normal epithelial cells introduced in a sampling process is avoided, it is ensured that the cells in which the epithelial cell marker genes and / or mesenchymal cell marker genes are detected are circulating tumor cells of breast cancer, and thus the accuracy and reliability of the detection result are further improved.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a breast cancer circulating tumor cell identification kit. Background technique [0002] Breast cancer is a malignant tumor that occurs in the glandular epithelial tissue of the breast, 99% of which occur in women, and only 1% in men. The mammary gland is not an important organ to maintain human life activities, and 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 off. Once the cancer cells fall off, the free cancer cells can spread throughout the body with the blood or lymph fluid, forming metastasis, which is life-threatening. At present, breast cancer has become a common tumor that threatens women's physical and mental health. Early detection and early diagnosis are the key to improving the curative effect. [0003] ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68
CPCC12Q1/6841C12Q1/6886C12Q2600/158C12Q2565/519C12Q2563/107
Inventor 刘苏燕吴诗扬董艳
Owner SUREXAM BIO TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap