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Circular RNA marker hsa_circ_0001788 and its application

A technology of markers and primer pairs, applied in preventive medicine, clinical medicine, and biotechnology, can solve the problem of less circRNA, achieve good stability, not easy to degrade, and high auxiliary diagnostic value

Active Publication Date: 2020-05-19
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, scholars at home and abroad have detected abnormally expressed circRNAs in the serum of patients with various diseases, but there are few reports on circRNAs related to craniosynostosis.

Method used

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  • Circular RNA marker hsa_circ_0001788 and its application
  • Circular RNA marker hsa_circ_0001788 and its application
  • Circular RNA marker hsa_circ_0001788 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 sample collection:

[0041] Twenty-six patients with newly diagnosed craniosynostosis and 23 matched controls were collected. All cases were confirmed by CT. Controls were patients with traumatic brain injury. The guardians of all research subjects signed the informed consent and provided 300 μL peripheral blood serum samples from the children.

Embodiment 2

[0042] Example 2 Serum RNA Extraction:

[0043]The operation was performed according to the instructions of the Invitrogen Trizol LS (10296028) kit. Specific steps are as follows:

[0044] ①Add 0.75mL TRIzol LS reagent to 0.25mL serum sample; ②Mix the sample, let stand at room temperature for 5min, centrifuge at 12000g at 4°C for 5min, and take the supernatant; ③Add 0.2mL chloroform, mix well, let stand at room temperature for 2-3min, 4°C Centrifuge at 12000g for 15min, take the supernatant; ④Add 0.5mL of isopropanol, place at room temperature for 10min, centrifuge at 12000g for 10min at 4℃, discard the supernatant and blot dry gently; ⑤Add 1mL of 75% ethanol, mix well, centrifuge at 4℃, 7500g, 5min; ⑥Discard the supernatant, invert on clean filter paper for a while, centrifuge at 5000g at 4°C for 3min; ⑦Remove ethanol with a pipette tip, add 20-50μL DEPC water, mix well; ⑧Water at 55-60°C for 10-15min, NanoDrop ND-2000 instrument detects RNA concentration and purity.

Embodiment 3

[0045] Example 3 circRNA expression profile screening

[0046] 5 cases of craniosynostosis patients and 5 cases of control population were randomly selected for peripheral blood total RNA, and Shanghai Kangcheng Bioengineering Co., Ltd. was commissioned to detect 5 cases of craniosynostosis patients and 5 cases of The expression profile of circRNAs in the peripheral blood serum of the control population was screened out to screen out the differentially expressed circRNA hsa_circ_0001788 for verification with expanded samples.

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Abstract

The invention establishes a circRNA expression profile for a patient with craniosynostosis, provides, provides serum circRNA marker hsa_circ_0001788 for the patient with craniosynostosis, discloses diagnostic value of hsa_circ_0001788 of serum for craniosynostosis, applies the marker herein to prepare a diagnostic kit for the patient with craniosynostosis, and provides supporting for clinical early discovery and treatment of the patient with craniosynostosis. Serum circRNA has good stability, degradation difficulty and the other advantage, has high value in auxiliary diagnosis, and is easy toclinically popularize and use. A circRNA chip is used herein to carry out detecting to obtain disease-specific and abnormally-expressed serum circRNA expression profiles; verifying is performed by means of qRT-PCR; a strict design and evaluation system is provided. It is discovered for the first time that the marker hsa_circ_0001788 in serum is an important biological detection index which causesminimal invasion and has low cost and which is applicable to clinical detection; the marker hsa_circ_0001788 is important to the clinical early diagnosis and differential diagnosis of craniosynostosis; a new idea is provided for the diagnosis of craniosynostosis in molecular level, and it is expected that early diagnostic rate of craniosynostosis is increased.

Description

technical field [0001] The invention belongs to the fields of biotechnology, preventive medicine and clinical medicine, and discloses a circRNA marker related to auxiliary diagnosis of craniosynostosis and its application, specifically, the circular RNA marker hsa_circ_0001788 and its application. Background technique [0002] Craniosynostosis is a common craniofacial deformity with an incidence rate of about 1 / 2500, ranking second in the incidence rate of congenital craniofacial bone deformities. It is characterized by premature fusion of one or more sutures before brain development is complete, resulting in restricted development of the skull, brain, face, and central nervous system. The disease will have a serious impact on the appearance of the patient. In some cases, the narrow cranial cavity can hinder the development of brain tissue, causing mental and motor development to lag behind. At present, the diagnosis of craniosynostosis is mainly based on abnormal head shap...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/166C12Q2600/178
Inventor 顾爱华王刚颜青徐进翁振坤刘倩
Owner NANJING MEDICAL UNIV
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