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Rapidly screened ovarian cancer nucleic acid aptamer and application thereof in preparation of preparation for detecting ovarian cancer

A nucleic acid aptamer, ovarian cancer technology, applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of difficult to detect pathological tissue, poor detection, etc., and achieve the effect of low cost, cost saving, and wide target range

Active Publication Date: 2020-11-17
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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  • Rapidly screened ovarian cancer nucleic acid aptamer and application thereof in preparation of preparation for detecting ovarian cancer
  • Rapidly screened ovarian cancer nucleic acid aptamer and application thereof in preparation of preparation for detecting ovarian cancer
  • Rapidly screened ovarian cancer nucleic acid aptamer and application thereof in preparation of preparation for detecting ovarian cancer

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 binding library isolated and retained 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 / 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 ...

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 / 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 wit...

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Abstract

The invention discloses a nucleic acid aptamer usable for detecting early ovarian cancer tissues and an application of the nucleic acid aptamer in early clinical detection of ovarian cancer. A 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 a traditional antibody, the nucleic acid aptamer has the advantages that the target recognition range is wide, the specificity is high, the affinity is high, the toxicity and immunogenicity are low, the in-vitro synthesis cost is low, the time is saved, theaptamer can be used for drug loading or modification, the functionality of the aptamer is enhanced, and the expected treatment target is achieved. By utilizing the short-chain nucleic acid aptamer andperforming modifying and marking, ovarian cancer epithelial cells and tissues can be identified and detected, the process is rapid, simple and convenient, and a new thought is provided for early detection and clinical treatment of the 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 ovary. 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, an...

Claims

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Application Information

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Patent Type & Authority Applications(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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