Parkinson diagnostic marker and application thereof

A technology for expressing products and genes, applied in the diagnostic markers of Parkinson's, the application field of FAM102A in the preparation of Parkinson's diagnosis and treatment reagents, can solve the problems that cannot meet the clinical needs, and achieve simplified quantitative detection process, clear results, fast response effect

Active Publication Date: 2015-11-18
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, genes related to hereditary PD have been found to include SNCA, LRRK2, DJ-1, PINK-1, Parkin, etc. These pathogenic genes need further research and are far from meeting clinical needs

Method used

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  • Parkinson diagnostic marker and application thereof
  • Parkinson diagnostic marker and application thereof
  • Parkinson diagnostic marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 High-throughput sequencing and analysis

[0029] The samples came from Peking Union Medical College Hospital, and the informed consent of the subjects was obtained. Collect peripheral blood samples from 15 cases of Parkinson's disease and 9 cases of healthy control peripheral blood samples for RNA extraction. After RNA extraction, agarose gel electrophoresis can be used to preliminarily determine whether the quality of the extracted RNA samples is qualified or not. for further transcriptome analysis. Then, the extraction of RNA samples was detected by NanoDrop1000 spectrophotometer, and the sample requirements for RNA-seq sequencing: OD260 / OD280 was 1.8-2.2.

[0030] The sequencing platform is Illumina's HiSeq2500 high-throughput sequencing platform, which performs high-throughput transcriptome deep sequencing. After sequencing, we use Fast-QC

[0031] (http: / / www.bioinformatics.babraham.ac.uk / projects / fastqc / ) The software evaluates the quality of sequencin...

Embodiment 2

[0032] Example 2 FAM102A gene expression in peripheral blood of patients with Parkinson's disease and peripheral blood of healthy people

[0033] 1. Materials and methods

[0034] 1. Materials

[0035] The peripheral blood of 135 cases of Parkinson's patients and 33 cases of healthy people were collected, grouped and numbered.

[0036] 2. Method

[0037] 2.1 Extraction of total RNA from the peripheral blood of Parkinson's patients and healthy people

[0038] The Kangwei Century Blood RNA Extraction Kit (Catalog No. CW0538) was used for sample RNA extraction, and the experimental operation was carried out according to the product manual. For specific operations, see the manual.

[0039] Judgment criteria for RNA quality: the OD260 / OD280 value of the RNA sample is between 1.7 and 2.2; the electrophoretic pattern of total RNA has clear 28S and 18S bands; difference.

[0040] 2.2 Design and synthesis of FAM102A detection primers

[0041] According to the principles of PCR pr...

Embodiment 3

[0071] Embodiment 3 A detection kit for detecting Parkinson's and its method of use

[0072] RNA extraction reagent: ultrapure RNA extraction kit (Cat. No. CW0597)

[0073] Fluorescent quantitative reagent: UltraSYBR one-step fluorescent quantitative PCR kit (Cat. No. CW0660)

[0074]

[0075]

[0076] Fluorescent quantitative PCR reaction system and method:

[0077] RT-PCR reaction system (25 μl): 2×UltraSYBROneStepRT-qPCRBuffer (WithROX) 12.5 μl, upstream primer (10 μM) 0.5 μl, downstream primer (10 μM) 0.5 μl, SuperEnzymeMix 0.5 μl, add RNA template (final concentration 10pg–100ng ), RNase-FreeWater to make up to 25 μl.

[0078] Reaction regulation: Reverse transcription: 10min at 45°C; pre-denaturation at 95°C for 10min, followed by 30-40 cycles: 15s at 95°C, 60s at 60°C.

[0079] The present invention uses high-throughput sequencing to screen out the Parkinson's disease-related gene FAM102A, combined with molecular biology experiment verification, it is confirmed...

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Abstract

The invention relates to a Parkinson diagnostic marker and application thereof, in particular to application of genes FAM102A to preparing Parkinson diagnosis and treatment reagents. The Parkinson diagnostic marker and the application have the advantages that genes are screened by an inventor on the basis of high-throughput sequencing results by the aid of bioinformatics methods and analysis, candidate genes FAM102A are selected, and relations between the genes FAM102A and Parkinson's diseases are proved by the aid of molecular biological methods; the genes FAM102A are in low-expression in the peripheral blood of Parkinson patients, have excellent correlation with Parkinson's diseases, can be used for preparing the auxiliary diagnosis and treatment reagents for the Parkinson's diseases and have important clinical application value.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular to a Parkinson's diagnostic marker and its application, more specifically to the application of FAM102A in the preparation of Parkinson's diagnosis and treatment reagents. Background technique [0002] Parkinson's disease (Parkinson's disease, PD) is a chronic progressive disease, currently incurable, it was first described by the British Parkinson in 1817, with the progress of PD research, we have a deeper understanding of this disease , it is clinically manifested as motor symptoms such as resting tremor, bradykinesia, muscle rigidity, and posture and gait disturbance and non-motor symptoms such as constipation, sleep disturbance, and abnormal mental behavior. At present, the pathogenesis of PD is still unclear, and it may be related to genetic factors, mitochondrial dysfunction, oxidative stress, immune abnormalities, cell apoptosis, and environmental factors. In PD, about 10%...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68A61K48/00A61K45/00A61K31/713A61P25/16
CPCA61K31/713A61K38/1709A61K45/00A61K48/00C12Q1/6883C12Q2600/158G01N33/68G01N2333/47G01N2800/2835
Inventor 杨承刚肖枫
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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