Application of TMF1 as Parkinsonism diagnosis marker
A Parkinson's disease and product technology, applied in the field of molecular diagnosis, can solve problems such as unstable posture, muscle stiffness, gait disorder, etc., and achieve more accurate genetic diagnosis, reduce mortality, and timely genetic diagnosis
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Embodiment 1
[0050] Example 1 Screening for genes differentially expressed in Parkinson's disease patients and normal individuals
[0051] 1. Research object:
[0052] Collect 10 cases of primary PD patients, 5 males and 5 females, aged 30-85 years, with a course of disease ranging from 3 months to 22 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.
[0053] Normal group: select 10 healthy volunteers aged 30-85, 5 males and 5 males.
[0054] There was no significant difference in age and gender between the two groups (P>0.10), which we...
Embodiment 2
[0080] Embodiment 2QPCR experiment verifies the genes differentially expressed in patients with Parkinson's disease and normal people
[0081] 1. Research object:
[0082] The screening criteria were the same as in Example 1, 50 cases of Parkinson's disease patients and 50 normal persons.
[0083] 2. Extraction of total RNA in blood
[0084] Step is with embodiment 1.
[0085] 3. Reverse transcription
[0086]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, and add the following components to the PCR tube: DEPC water, 5× reverse transcription buffer, 10mmol / LdNTP, 0.1mmol / lDTT, 30μmmol / lOligodT, 200U / μl M-MLV, template RNA. Incubate at 42°C for 1 hour, then centrifuge briefly at 72°C for 10 minutes.
[0087] 4. QPCR
[0088] (1) Primer design
[0089] QPCR amplification primers were designed according to the coding sequences of TMF1 gene and GA...
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