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Gene, detection method and kit used for detecting depressive disorder

A detection kit and technology for depression, applied in the field of molecular biology, can solve the problems of lack of early prevention and lack of diagnosis, and achieve the effects of strong specificity, high sensitivity and accurate detection

Active Publication Date: 2014-02-12
BEIJING CENT FOR PHYSICAL & CHEM ANALYSIS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the persistent psychological disorder appears for at least two weeks, and the severe state can only be confirmed for at least two years, the current diagnosis lacks rapid and sensitive indicators, and it is difficult to confirm whether the patient has the disease or has developed it in a short period of time, let alone measures. Early prevention to avoid tragedy

Method used

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  • Gene, detection method and kit used for detecting depressive disorder
  • Gene, detection method and kit used for detecting depressive disorder
  • Gene, detection method and kit used for detecting depressive disorder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1A

[0045] Example 1 Amplification of specific fragments of ATP1A1 gene

[0046] With the consent of the blood donors, a large amount of blood was collected from normal people and patients with clinically diagnosed depression with anticoagulation tubes, and the blood collection tubes were immediately placed in an ice box. All blood samples were analyzed by molecular biology method, compared with the human genome sequences reported in the NCBI database, and the relative expression levels of the same gene in different individual samples were calculated using the fluorescence quantitative PCR method according to the standard curve. The gene expression level of the same gene and the normal person were statistically compared and analyzed. The expression level of the depression patients and the normal person was significantly different, which was considered to be a depression-specific related gene. The results were found in the NCBI database serial number 476. The expression of ATP1A1 g...

Embodiment 2

[0058] Example 2 Establishment of a real-time fluorescent quantitative PCR method for detecting ATP1A1 gene

[0059] Using the cDNA after reverse transcription of total RNA in blood as a template, and using the specific primers involved in Example 1 as PCR reaction primers, a real-time fluorescence quantitative PCR reaction method for detecting ATP1A1 gene was established. This embodiment adopts Fast Green Master Mix fluorescent dye (applied biology company, product number 4385612) real-time quantitative PCR reaction 20μl The reaction system is: template 2μl, 10mmol upstream primer 0.5μl, 10mmol downstream primer 0.5μl, fast SYBR-Green Master Mix 10μl, deionized water 7μl .

[0060] Reaction conditions: pre-denaturation at 95°C for 20s, (95°C for 3s, 60°C for 30s) for 40 cycles.

[0061] Dissolution curve analysis: 95℃ for 15s, 60℃ for 1min, 95℃ for 15s, 60℃ for 15s.

[0062] The standard quantitative expression curve of normal control gene and depression-specific ATP1A1 g...

Embodiment 3

[0063] Example 3 Clinical application experiment 1

[0064] Venous blood from patients with depression was collected, collected in an anticoagulant tube, and immediately put into an ice box, and the test was started 4 hours later. with PureLink TM RNA Mini Kit (Invitrogen Item No. 12183018A) was used to extract RNA from blood, and the RNA concentration was determined to be 32.03 ng / ul. After using the fluorescence quantitative PCR of Example 5 of the present invention, the gene ATP1A1 in the venous blood of patients with depression was calculated as 33 copies, lower than normal (9447 copies for normal)

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Abstract

The invention provides a target gene and a detection kit used for detecting depressive disorder. The target gene used for detecting the depressive disorder is ATP1A1, and nucleotide sequences of a real-time fluorescence quantification PCR (polymerase chain reaction) primer for detecting the gene are shown as SEQ ID NO.2-3. The invention also provides the detection kit used for quantitative detection on ATP1A1. Effective guidance is provided for clinic detection of the depressive disorder by application of the ATP1A1 in preparation of medicines for diagnosing, treating and preventing the depressive disorder.

Description

technical field [0001] The present invention relates to the field of molecular biology, in particular to the target gene ATP1A1 for detecting depression, and the present invention also relates to a method and kit for detecting the ATP1A1 gene. Background technique [0002] Depression is a common mental illness with mood disorders as the main manifestation. Its specific etiology is still unclear. The main clinical features are marked and persistent low mood, and the low mood is not commensurate with the environment in which they live. In severe cases, suicide may occur. thoughts and actions. At present, depression has become the fourth largest disease in the world. It is expected that by 2020, it may become the second largest human disease after heart disease. Depression will become the main killer of human beings in the 21st century. [0003] At present, the diagnosis of depression is still limited to the diagnosis of clinical symptoms. Due to the persistent psychological ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/11C12Q1/68G01N21/64
Inventor 武会娟吴宁李丽李越钱嘉林李宝明张小莉刘旭
Owner BEIJING CENT FOR PHYSICAL & CHEM ANALYSIS
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