Biomarker for detecting mild cognitive impairment and/or Alzheimer disease

A technology for Alzheimer's disease and mild cognitive impairment, applied in the field of biomarkers for the detection of mild cognitive impairment and/or Alzheimer's disease, can solve the problem of non-specific detection methods and mild cognitive impairment Obstacles such as limited blood test markers

Inactive Publication Date: 2018-06-22
SHENZHEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, provide a kind of mild cognitive impairment and/or Alzheimer's disease biomarker that can be detected by peripheral blood samples, and be applied to the detection of human blood sampl

Method used

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  • Biomarker for detecting mild cognitive impairment and/or Alzheimer disease
  • Biomarker for detecting mild cognitive impairment and/or Alzheimer disease
  • Biomarker for detecting mild cognitive impairment and/or Alzheimer disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The detection of ciRS-7 comprises the following steps:

[0035] 1. Primer design for circRNA biomarker (i.e. ciRS-7)

[0036] In this experiment, the amplification primers of circRNA biomarkers and internal reference gene (GAPDH) were first designed, and the specific sequences are shown in Table 1.

[0037] Table 1

[0038]

[0039]

[0040] 2. RNA extraction

[0041] Use the blood RNA extraction kit to extract RNA from human peripheral blood (whole blood, serum, plasma, white blood cells), and then perform RNA quality detection. Ultra-micro spectrophotometer detection, A260 / A280 > 1.8; electrophoresis detection, if it is not degraded, the RNA quality is qualified.

[0042] 3. Reverse transcription

[0043] 3.1 Genomic DNA removal reaction

[0044] Prepare the reaction mixture on ice according to the composition in Table 2 below, and carry out the DNA removal reaction. Reaction conditions: put the reaction mixture prepared in Table 2 at 42°C for 2 minutes. ...

Embodiment 2

[0061] Using the ciRS-7 detection steps constructed in Example 1, apply:

[0062] 1. Detection of ciRS-7 (CDR1as) in whole blood, serum / plasma, and white blood cells

[0063] A. RNA was extracted from 200 μL whole blood of normal people (25 years old and 52 years old), reverse-transcribed into cDNA and then tested by qPCR. The results are as follows figure 1 and figure 2 shown. From figure 1 It can be seen that the melting curve is a single peak and the Tm value is greater than 80°C, excluding the possibility of non-specific amplification and primer dimer, indicating that ciRS-7 is contained in whole blood. From figure 2 It can be seen that the content of ciRS-7 in whole blood increases with age.

[0064] B. Extract RNA from leukocytes obtained from 2.5ml whole blood of normal people (25 years old and 52 years old), reverse transcribe into cDNA for qPCR detection, the results are as follows image 3 and Figure 4 shown. From image 3 It can be seen that the dissolut...

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Abstract

The invention belongs to the technical field of biological detection, and particularly relates to a biomarker for detecting mild cognitive impairment (MCI) and/or Alzheimer disease (AD). The biomarkerfor detecting mild cognitive impairment and/or Alzheimer disease is circular RNA, and the circular RNA is ciRS-7. The invention further relates to application of the circular RNA ciRS-7 in detectingmild cognitive impairment and/or Alzheimer disease. By detecting the content of the ciRS-7 in peripheral blood, it is found that the content of the ciRS-7 in the peripheral blood of an MCI or AD patient is higher than that of a normal person, and therefore the ciRS-7 can be used as the biomarker of MCI or AD early diagnosis.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a biomarker for detection of mild cognitive impairment and / or Alzheimer's disease. Background technique [0002] At present, the main methods for clinical diagnosis of mild cognitive impairment (MCI) and Alzheimer's disease (Alzheimer disease, AD) include scale detection, imaging detection, and cerebrospinal fluid biomarker detection. Scale testing is not accurate enough due to individual subjectivity; imaging testing requires expensive equipment such as MRI and PET, which cannot be widely used; cerebrospinal fluid sampling is invasive and difficult, and patients and their families are often reluctant to cooperate. Currently there is no high accuracy and specific peripheral blood biomarker for MCI and AD. [0003] Circular RNA (circRNA) is a new type of RNA that is different from traditional linear RNA. It has a closed circular structure and exists in la...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 吴冲刘琼
Owner SHENZHEN UNIV
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