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Combinations of miRNA markers, kits and methods for detecting breast cancer

A marker, breast cancer technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc.

Active Publication Date: 2021-12-24
SHANGHAI SANTEJA BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a need to identify specific microRNA markers or combinations of microRNA markers to detect breast cancer more efficiently

Method used

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  • Combinations of miRNA markers, kits and methods for detecting breast cancer
  • Combinations of miRNA markers, kits and methods for detecting breast cancer
  • Combinations of miRNA markers, kits and methods for detecting breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099]

[0100] Five microRNAs (miR-21, miR-29a, miR-101, miR-221, and miR-155) implicated in association with breast cancer were evaluated in detail by the training group[ figure 1 ]. In the training set, the improvement of the marker properties achieved by using the most effective markers of these five microRNAs and their combinations was discussed, and thus the analysis using the bootstrapping method was performed. Quantitative analysis of each microRNA used the quantitative PCR method by reverse transcription reaction. The details of the experiment are described below.

[0101] plasma separation : Use a 5mL or 7mL capacity EDTA-2Na vacuum blood collection tube to collect venous blood from breast cancer patients and healthy subjects, and immediately use it for centrifugation (2500×g / 10 minutes) at room temperature after inversion mixing. Then, the upper plasma fraction was dispensed into another tube with a disposable pipette, and stored at -80°C until use.

[0102] ...

Embodiment 2

[0150]

[0151] With respect to the evaluation markers performed in the training group of Example 1, the discrimination accuracy of each marker was evaluated based on AUC using logistic regression analysis for a validation group consisting of another cohort. Here, miR-101 and miR-221 were selected from the evaluation markers performed in the training group, and the discrimination accuracy of each marker was evaluated based on AUC using logistic regression analysis. Regarding the experimental method, according to Example 2, the microRNA in the purified plasma was used to carry out reverse transcription reaction and quantitative PCR method. Details are described below.

[0152] plasma separation : Use a 5mL or 7mL capacity EDTA-2Na vacuum blood collection tube to collect venous blood from breast cancer patients and healthy subjects, and immediately use it for centrifugation (2500×g / 10 minutes) at room temperature after inversion mixing. Then, the upper plasma fraction was d...

Embodiment 3

[0183]

[0184] With respect to the evaluation markers performed in the training group of Example 1, the discrimination accuracy of each marker was evaluated based on AUC using logistic regression analysis for a validation group consisting of another cohort. Here, miR-101 and miR-155 were selected from the evaluation markers performed in the training group, and the discrimination accuracy of each marker was evaluated based on AUC using logistic regression analysis. Regarding the experimental method, according to Example 3, the microRNA in the purified plasma was used to carry out reverse transcription reaction and quantitative PCR method. Details are described below.

[0185] plasma separation : Use a 5mL or 7mL capacity EDTA-2Na vacuum blood collection tube to collect venous blood from breast cancer patients and healthy subjects, and immediately use it for centrifugation (2500×g / 10 minutes) at room temperature after inversion mixing. Then, the upper plasma fraction was d...

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Abstract

The present invention provides a biomarker technology for diagnosing the mammary gland of an examinee with high specificity and sensitivity by measuring the expression levels of two or more miRNAs circulating in human peripheral blood in combination cancer. Regardless of the type of breast cancer, the clinical stage, and the differences in race, common miRNA markers can be used, and they all have high detection sensitivity and specificity. These breast cancer markers are miRNAs confirmed from multiple cohort studies, and are techniques that can be directly used in clinical examinations as breast cancer biomarkers.

Description

technical field [0001] The present application relates to a biomarker technique for examining breast cancer using serum or plasma collected from a test subject. Background technique [0002] Breast cancer is one of the major diseases threatening women's health. According to the statistics of the World Health Organization (WHO: World Health Organization) in 2008, breast cancer ranks the fifth cause of death of adult women in the world. Breast cancer is broadly divided into four types according to its genotype: Luminal A type, Luminal B type, HER2Rich type, and Triple Negative type. It is known that the sensitivity to anticancer agents is a difference in the nature and malignancy of cancer cells. In Luminal A and B types, estrogen receptor (Estrogen Receptor; ER) and progesterone receptor (Progesterone Receptor; PgR) are positive, and in Luminal B type and HER2 Rich type, HER2 receptor is positive. In addition, when viewed from the degree of progression of cancer invasion, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 落谷孝広
Owner SHANGHAI SANTEJA BIOTECH CO LTD
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