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Diagnostic use of the nacc2 gene in blood

A gene and gene expression technology, applied in the field of molecular diagnosis, to achieve the effect of reducing mortality and timely gene diagnosis

Active Publication Date: 2018-11-30
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The complexity of PD is also reflected in the clinical challenges
To date, no diagnostic test exists for definitive diagnosis of early stages of PD

Method used

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  • Diagnostic use of the nacc2 gene in blood
  • Diagnostic use of the nacc2 gene in blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Screening for genes differentially expressed in Parkinson's disease patients and normal individuals

[0036] 1. Research object:

[0037] Collect 10 cases of primary PD patients, 5 males and 5 females, aged 32-85 years, with a course of disease ranging from 5 months to 20 years. PD inclusion criteria: all diagnostic criteria meet the clinical diagnostic criteria for PD (refer to "Jiang Yuping, Wang Jian, Ding Zhengtong, et al., Diagnostic criteria for primary Parkinson's disease, 2005, Chinese Clinical Neuroscience, 2006, 14:40" ). Exclusion criteria: (1) essential tremor; (2) secondary parkinsonism; (3) severe dementia, dysarthria; (4) patients with other mental diseases. This study was approved by the Hospital Ethics Committee and all patients signed informed consent.

[0038] Normal group: 10 healthy volunteers aged 32-80 were selected, 5 males and 5 males.

[0039] There was no significant difference in age and gender between the two groups (P>0.10), w...

Embodiment 2

[0065] Example 2 QPCR experiment verification of genes differentially expressed in patients with Parkinson's disease and normal people

[0066] 1. Research object:

[0067] The screening criteria were the same as in Example 1, 50 cases of Parkinson's disease patients and 50 normal persons.

[0068] 2. Extraction of total RNA in blood

[0069] Step is with embodiment 1.

[0070] 3. Reverse transcription

[0071] 1 μg of total RNA was reverse-transcribed to synthesize cDNA using reverse transcription buffer. Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, add the following components to the PCR tube respectively: DEPC water, 5× reverse transcription buffer, 10mmol / L dNTP, 0.1mmol / l DTT, 30μmmol / l Oligo dT, 200 U / μl M-MLV, template RNA. Incubate at 42°C for 1h, then centrifuge briefly at 72°C for 10min.

[0072] 4. QPCR

[0073] (1) Primer design

[0074] QPCR amplification primers were designed according to the coding sequences of NACC2 gene...

Embodiment 3

[0090] Example 3 Verification of expression products of differentially expressed genes in patients with Parkinson's disease and normal people by immunoblotting experiments

[0091] 1. Clinical object: same as embodiment 2.

[0092] 2. Mononuclear cell isolation

[0093] Take 10ml of venous blood from patients with Parkinson's disease and normal people, inject it into a sterile vial containing heparin, and shake it gently immediately after capping. Add an equal volume of HBSS (NaCl 8.0g, NaCl 2 HPO 4 0.132g, KH 2 PO 4 0.06g, KCl0.4g, phenol red 1ml, NaHCO 3 0.35g, D-glucose 1.0g, dissolved in 1000ml double distilled water), to reduce the aggregation of red blood cells. Draw 8ml of lymphocyte stratification solution into a 50ml centrifuge tube, slowly add the diluted blood along the tube wall, keep the interface clear, do not mix the two, centrifuge at 20°C 2000r / min for 30min, carefully absorb the stratification solution and plasma transfer The turbid off-white layer,...

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Abstract

The invention discloses a diagnostic tool for the Parkinson's disease. The diagnostic tool is used for realizing a diagnostic purpose through detecting an NACC2 gene and an expression product thereofin blood. Proven by the search of the diagnostic tool, in comparison with a normal person, the expression level of mRNA (messenger Ribonucleic Acid) of the NACC2 gene in the blood of a patient suffered from the Parkinson's disease is obviously raised. According to the relevance existing between the NACC2 gene and the Parkinson's disease, a kit for diagnosing the Parkinson's disease can be prepared, and the kit can be clinically widely applied.

Description

technical field [0001] The invention belongs to the field of molecular diagnosis, and relates to a molecular marker for the diagnosis of Parkinson's disease, in particular to the application of the molecular marker-NACC2 gene in blood in the preparation of products for diagnosing Parkinson's disease. Background technique [0002] Parkinson's disease (PD) is a common neurodegenerative disease that mainly occurs in middle-aged and elderly people. With the intensification of global aging, the number of patients with Parkinson's disease continues to expand. It not only seriously affects the quality of life of patients and their families, but also brings an increasingly heavy economic burden to society. [0003] As the most common neurodegenerative movement disorder, Parkinson's disease (PD) is characterized by several motor symptoms, such as resting tremor, muscle rigidity, and postural instability, as well as some non-motor symptoms, such as autonomic disturbance, Mental abnor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6837C12N15/11G01N33/68
CPCC12Q1/6837C12Q1/6883C12Q2600/158G01N33/6896
Inventor 汪冰怡肖枫
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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