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A rapidly screened ovarian cancer nucleic acid aptamer and its application in the preparation and detection of ovarian cancer preparations

A technology of nucleic acid aptamer and ovarian cancer, applied in measuring devices, biological testing, material inspection products, etc., can solve the problems of difficult detection of pathological tissue and poor detection, and achieve low cost, time-saving and low cytotoxicity

Active Publication Date: 2022-03-11
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Imaging diagnosis includes CT, MRI (magnetic resonance imaging), PET, etc., and CT is less than 2cm, MRI is poor in detecting small lesions, and PET is difficult to detect pathological tissues less than 1cm

Method used

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  • A rapidly screened ovarian cancer nucleic acid aptamer and its application in the preparation and detection of ovarian cancer preparations
  • A rapidly screened ovarian cancer nucleic acid aptamer and its application in the preparation and detection of ovarian cancer preparations
  • A rapidly screened ovarian cancer nucleic acid aptamer and its application in the preparation and detection of ovarian cancer preparations

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Nucleic acid aptamer screening of clinical ovarian cancer tissue

[0027] Based on Tissue-SELEX technology, positive screening is performed on clinical ovarian cancer tissues, and negative screening is performed on ovarian cancer paracancerous tissues to obtain the expected target sequence. Synthesize the X-Aptamer library with affinity group modification first. In the first round of screening, the initial library was incubated with ovarian cancer paracancerous tissues, and the isolated and retained binding library was the negative screening library. The non-binding library was incubated with ovarian cancer tissues, and the binding library that was isolated and retained was a positive screening library. In the second round of screening, the positive screening library and negative screening library in the first round of screening were mixed with sterile deionized water (blank group), ovarian cancer tissue (positive group), and ovarian cancer adjacent tissue (...

Embodiment 2

[0031] Example 2: Determining the sequence with the strongest specific binding ability to the HO-8910 cell line

[0032] Centrifuge the powder of the 8 nucleic acid aptamer sequences and the control library chain, dilute it with sterile deionized water to a final concentration of 250nM, denature it in a metal bath at 95°C for 5min, refold it on ice for 10min, and place it at room temperature. At the same time, the HO-8910 adherent cells were washed twice with DPBS, and 0.2% EDTA was added to digest the cells at 37°C. After washing twice with DPBS, the cells were collected by pipetting with washing buffer (Washing buffer, containing 0.45% glucose, 5 mM magnesium chloride) into a centrifuge tube. After centrifugation and washing, add binding buffer (Binding Buffer: DPBS, 0.45% glucose, 5 mM magnesium chloride, 100mg / L tRNA, 1g / L BSA) and the processed nucleic acid aptamer sequence and library. Mix by pipetting and incubate on a shaker at 4°C for 1 hour. After incubation, wash...

Embodiment 3

[0035] Example 3: Determining the sequence with the strongest specific binding ability to the A2780 cell line

[0036] Centrifuge the powder of the 8 nucleic acid aptamer sequences and the control library chain, dilute it with sterile deionized water to a final concentration of 250nM, denature it in a metal bath at 95°C for 5min, refold it on ice for 10min, and place it at room temperature. At the same time, the A2780 adherent cells were washed twice with DPBS, and 0.2% EDTA was added to digest the cells at 37°C. After washing twice with DPBS, the cells were collected by pipetting with washing buffer (Washing buffer, containing 0.45% glucose, 5 mM magnesium chloride) into a centrifuge tube. After centrifugation and washing, add binding buffer (Binding Buffer: DPBS, 0.45% glucose, 5 mM magnesium chloride, 100mg / LtRNA, 1g / L BSA) and the processed nucleic acid aptamer sequence and library. Mix by pipetting and incubate on a shaker at 4°C for 1 hour. After incubation, wash with...

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Abstract

The invention discloses a nucleic acid aptamer that can be used to detect early ovarian cancer tissue and its application in early clinical detection of ovarian cancer. The sequence of the nucleic acid aptamer is selected from any one of SEQ ID NO.1 to SEQ ID NO.8, or the sequence is optimized or modified. Compared with traditional antibodies, the nucleic acid aptamer in the present invention has a wide range of recognized targets and strong specificity, strong affinity, low toxicity and immunogenicity, low cost and time-saving in vitro synthesis, and can be loaded with aptamers Or modified to enhance the functionality of the aptamer and achieve the desired therapeutic goals. Utilizing the short-chain nucleic acid aptamer of the present invention and modifying and marking it, the epithelial cells and tissues of ovarian cancer can be identified and detected, and the process is fast and simple, providing a new idea for early detection and clinical treatment of ovarian cancer.

Description

technical field [0001] The invention relates to a nucleic acid aptamer and its application, in particular to a nucleic acid aptamer which can be used for detection of ovarian cancer epithelial cells and early clinical ovarian cancer tissue samples and its application in the preparation of detection reagents. Background technique [0002] Ovarian cancer is cancer caused by tumors in the ovaries. It is also the female reproductive system malignancy with the worst prognosis and a gynecological malignancy with low incidence and high mortality. [0003] According to the 2019 V3 version of the NCCN Clinical Practice Guidelines for Ovarian Cancer, ovarian cancer includes high-grade serous carcinoma, low-grade serous carcinoma, mucinous carcinoma, endometrioid carcinoma, and clear cell carcinoma. Type I ovarian cancers include endometrioid, clear cell, low-grade serous, and mucinous carcinomas. Type II ovarian cancer mainly consists of high-grade serous carcinoma, carcinosarcoma, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115G01N33/574G01N33/53
CPCC12N15/115G01N33/57449G01N33/5308C12N2310/16
Inventor 胡小晓谭蔚泓刘乃瑜文超琪
Owner HUNAN UNIV
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