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Application of exosomal mRNA in the diagnosis of breast diseases

An exosome, breast cancer technology, applied in the field of biomedical diagnosis, can solve the problems of inapplicable clinical disease diagnosis, unsuitable clinical diagnosis, different problems, etc.

Active Publication Date: 2021-06-25
JIANGSU MICRODIAG BIOMEDICINE TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the breasts of Chinese women are small and dense, which will also reduce the positive rate of patients receiving mammography
[0007] 3. Early detection of biological targets: including general screening of breast cancer-related genes, such as BRCA1 and BRCA2 mutation analysis, but current molecular diagnosis research mostly focuses on the relationship between gene polymorphisms and breast cancer, which can only give the susceptibility risk of patients, not Applicable to the diagnosis of clinical diseases
In addition, the most commonly used serum tumor markers for breast cancer in clinical practice include CA-153, CEA, CA-125, etc. Although they can reflect the state of the disease to a certain extent, the specificity is not high. In many other tumors, such as ovarian cancer, Pelvic tumors, gastric cancer, colon cancer, liver cancer, etc. will exist accordingly
It is reported that the sensitivity of CA-153 is only about 50%, and the expression level in early breast cancer patients is often low, and the specificity is not high, which has certain limitations in the early screening of breast cancer
[0008] 4. Needle biopsy: The pathological examination of breast tissue needle biopsy has high accuracy, but it is an invasive operation, and false negative results may be produced due to insufficient specimen collection, or due to different sampling sites, intratumoral abnormalities in breast cancer, etc. Qualitative effects can produce different results
[0009] In general: the screening of breast nodules has the following deficiencies: (1) the cost of clinical physical examination is high; (2) the rate of positive and missed detection of imaging diagnostic methods is high; (3) currently more mature biological targets, only Can do risk assessment, not suitable for clinical diagnosis; (4) Puncture biopsy is firstly invasive, and has high requirements for the sampler and puncture site, and it is easy to cause different results due to the influence of external factors

Method used

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  • Application of exosomal mRNA in the diagnosis of breast diseases
  • Application of exosomal mRNA in the diagnosis of breast diseases
  • Application of exosomal mRNA in the diagnosis of breast diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0190] In the following examples, it is analyzed by ENAH mRNA assay, which is designed to run on a fluorescent quantitative PCR detection platform to detect the expression level of ENAH mRNA in tissue exosomes and plasma exosomes. Using Lightcycle480 platform detection.

[0191] The experimental method is as follows:

[0192] 1. Plasma Processing:

[0193] Whole blood was collected using EDTA, and plasma was obtained by centrifugation within 2 hours at room temperature. Centrifugation conditions: 1500g, 10 minutes, absorb the supernatant, centrifuge again: 15000g, centrifuge for 10 minutes, absorb the supernatant, which is the separated plasma.

[0194] 2. Exosome identification:

[0195]2 mL of plasma was filtered with a 0.2 μm filter membrane, exosomes were enriched with a QIAGEN kit (exo-RNeasy Serum / Plasma Midi Kit), and the exosomes were eluted with the exosome eluent to obtain the structure Intact exosomes. Extracted exosomes were identified by particle tracking ana...

Embodiment 2

[0208] The type of sample tested in this embodiment is: plasma. A total of 39 patients were enrolled, and imaging tests showed that nodules existed in all of them. 19 samples of malignant breast nodules, specifically 11 cases of ductal carcinoma in situ with microinvasion, 7 cases of ductal carcinoma in situ, and 1 case of lobular carcinoma in situ; 20 cases of benign breast nodules, specifically 13 cases of fibroadenoma, 5 cases of There were 2 cases of papilloma and 2 cases of ductal epithelial hyperplasia.

[0209] See Example 1 for plasma exosome extraction, RNA extraction and detection. The test results are shown in Table 3.

[0210] Table 3: Differentiation between benign and malignant breast nodules

[0211]

[0212]

[0213] Results: Among 19 cases of malignant breast nodules, 14 cases were interpreted as positive, and the sensitivity was 73.68%; among 20 cases of benign breast nodules, 18 cases were interpreted as negative, with a specificity of 90%.

[0214...

Embodiment 3

[0216] The test sample type in this embodiment is: plasma, and a total of 91 samples were enrolled. Among them, 37 cases of breast cancer: 3 cases of intermediate-grade ductal carcinoma in situ, 5 cases of high-grade ductal carcinoma in situ, 17 cases of invasive ductal carcinoma, 3 cases of lobular carcinoma in situ, and 9 cases of invasive lobular carcinoma; 54 cases of non-breast cancer: 13 cases of lung cancer, 10 cases of colorectal cancer, 13 cases of benign breast disease (7 cases of mastitis, 4 cases of papilloma, 2 cases of ductal epithelial hyperplasia) and 18 cases of healthy people.

[0217] See Example 1 for plasma exosome extraction, RNA extraction and detection. The test results are shown in Table 4.

[0218] Table 4: Auxiliary diagnosis of breast cancer

[0219]

[0220]

[0221]

[0222] Results: 37 cases of breast cancer, 28 cases were positive, the sensitivity was: 75.68%; 54 cases of non-breast cancer, 47 cases were negative, the specificity was:...

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Abstract

The present invention relates to the field of biomedical diagnosis, in particular to the application of exosome mRNA in the diagnosis of breast diseases. The invention can distinguish benign from malignant breast nodules, effectively distinguish breast cancer from other malignant tumors, realize accurate diagnosis of breast cancer, and monitor the treatment effect of patients. The present invention finds for the first time that exosome mRNA markers can be used for the diagnosis of breast diseases.

Description

technical field [0001] The present invention relates to the field of biomedical diagnosis, in particular to the application of exosome mRNA in the diagnosis of breast diseases. Background technique [0002] Breast cancer (mammary cancer) is one of the most common malignant tumors in women. The incidence rate accounts for 7-10% of all kinds of malignant tumors in the whole body. It is second only to uterine cancer in women and has become the main cause of threat to women's health. Its incidence is often related to heredity, and between the ages of 40 and 60, the incidence rate of women before and after menopause is higher. It is one of the most common malignant tumors that usually occurs in the glandular epithelial tissue of the breast, seriously affecting women's physical and mental health and even threatening their lives. Breast cancer is rare in men, and only about 1-2% of breast cancer patients are men. [0003] In recent years, the incidence of breast cancer in my coun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 巴兆粉丁凤英韩春苗王弢
Owner JIANGSU MICRODIAG BIOMEDICINE TECH CO LTD
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