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Application of miRNA410- in preparation of prostatic cancer diagnostic kit

A mirna410-, diagnostic kit technology, applied in the field of gene therapy and medical diagnosis, can solve the problem of inability to further translate

Inactive Publication Date: 2015-05-27
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

miRNAs come from some RNAs that are transcribed from DNA but cannot be further translated into proteins (non-coding RNAs)
There is no literature report on the function research of human miRNA410-

Method used

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  • Application of miRNA410- in preparation of prostatic cancer diagnostic kit
  • Application of miRNA410- in preparation of prostatic cancer diagnostic kit
  • Application of miRNA410- in preparation of prostatic cancer diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] A total of 327 cases of human peripheral blood samples were detected, including 57 cases of normal subjects, 149 cases of prostate cancer, and the rest were benign prostatic hyperplasia. Among the prostate cancer specimens, 67 cases were mild prostate cancer (Gleason score less than 7), and 82 cases were severe prostate cancer (Gleason score greater than 7). The above samples were all provided by Changhai Hospital Affiliated to the Second Military Medical University of the Chinese People's Liberation Army.

[0086] Detection method: take a human peripheral blood sample, put it into a coagulation-promoting tube and centrifuge to extract serum, then use Trizol one-step method to extract serum RNA, and perform reverse transcription of miRNA according to steps 1 and 2 above. Then follow step 4 to detect the content of miR-410-.

[0087] Test results: Serum miR-410- in these specimens was detected, and it was found that a large amount of miR-410- could be detected in prosta...

Embodiment 2

[0096] The process of clinically detecting the content of serum miR-410- to assist in judging prostate cancer or normal population is as follows: figure 1 Shown:

[0097] figure 1 A: Obtain the peripheral blood of the patient to be tested;

[0098] figure 1 B: Obtain serum by centrifugation;

[0099] figure 1 C: Using Trizol one-step method to obtain total RNA from peripheral blood;

[0100] figure 1 D: total RNA of peripheral blood;

[0101] figure 1 E: miRNA is reverse transcribed into cDNA by tailing linker method;

[0102] figure 1 F: cDNA was amplified with miR410-specific primers;

[0103] figure 1 G: Analyze the amplification results using the standard curve.

[0104] The specific clinical applications are as follows:

[0105] Three patients were detected, and the miR-410- content was detected by the following method, combined with the clinical PSA content and the conclusion of pathological puncture, to determine whether they were suffering from prostate ca...

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PUM

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Abstract

The invention relates to the field of gene therapy and medical diagnosis, and provides application of miRNA410- in preparation of a prostatic cancer diagnostic kit. The invention further provides a kit for carrying out prostatic cancer diagnosis by utilizing quantitative PCR (Polymerase Chain Reaction) detection aiming at miR-410-. The application comprises the following steps: (1), after extracting total RNA of serum by using a phenol-chloroform method, carrying out reverse transcription of miRNA by utilizing a specific primer, so that miRNA is connected to a specific primer joint while reverse transcription is carried out; (2), carrying out quantitative PCR detection of cDNA by using a specific primer of miR-410-, analyzing by using a special standard curve, and calculating the content of serum miR-410-; and (3), diagnosing prostatic cancer by utilizing the content of serum miR-410-. The kit and the detection technology disclosed by the invention are mature, short in period and good in specificity.

Description

technical field [0001] The invention relates to the fields of gene therapy and medical diagnosis, in particular to the application of miRNA410-in the preparation of a prostate cancer diagnostic kit. Background technique [0002] Prostate cancer is a malignant tumor that occurs in male prostate tissue and is the result of abnormal and disordered growth of prostate acinar cells. In developed countries and regions such as Europe and the United States, it is the most common malignant tumor in men, and its mortality rate ranks second among various cancers; in Asia, its incidence rate is lower than that in Western countries, but it has shown a rapid upward trend in recent years. In 2007, the incidence of prostate cancer among malignant tumors of the genitourinary system in Shanghai jumped from No. 3 to No. 1, and quickly rose from No. 9 to No. 5 among the top ten male tumors. It is predicted that 10 years later, the incidence rate of prostate cancer in Shanghai will be promoted t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/178
Inventor 孙颖浩刘善荣张丹丹汪佳祺胡晶晶刘淑鹏肖潇程凯
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY