Process of detecting cell splitting periodic protein gene 6 in tumor using modified primer

A cell division and cycle protein technology, applied in the field of detection of cell division cycle protein gene 6 in tumors, can solve problems such as unreliable measurement results, achieve the effects of improving detection sensitivity and specificity, preventing infection, and ensuring reliability

Inactive Publication Date: 2007-07-18
张曼
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Problems solved by technology

However, precisely because the PCR reaction depends on high specificity and sensitivity, unexpected mistakes often occur in the determination of cdc6, resulting in unreliable determination results

Method used

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  • Process of detecting cell splitting periodic protein gene 6 in tumor using modified primer
  • Process of detecting cell splitting periodic protein gene 6 in tumor using modified primer

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Embodiment Construction

[0010] 1. Randomly select cases

[0011] 30 cases of bladder cancer were diagnosed, including 6 cases of initial onset and 24 cases of recurrence; 26 cases of men aged 36 to 84; 4 cases of women aged 60 to 73.

[0012] 25 cases of other urinary system diseases were diagnosed, including 18 cases of males aged 28-77; 7 cases of females aged 27-69; causes: 4 cases of bladder stones; 4 cases of kidney stones; 10 cases of urinary tract infection; 2 cases of kidney cancer; epididymitis 1 case; 2 cases of benign prostatic hyperplasia; 2 cases of urethral stricture. 5 male healthy volunteers aged 29-31.

[0013] 2. Preparation of urine exfoliated cells

[0014] Take 150-200ml of random urine after the first urine in the morning, divide it into several treated glass centrifuge tubes, centrifuge at 4°C and 380g for 10 minutes, discard the supernatant, collect the precipitate in a centrifuge tube, and wash with normal saline The pellet was washed twice, and the pellet was divided into...

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Abstract

The present invention relates to improved primer process for detecting cell division cycle protein gene 6 in tumor. In the process of DNA extraction, RNA extraction, protein extraction, immunohistochemistry, reverse transcription (RT), PCR, Northern hybridizing, in-situ hybridizing and capillary electrophoresis, is adopted an improved primer, which has the hairpin structure formed with the downstream primer 5' end base sequence and the primer 3' end base in the first reaction. At normal temperature, the primer 3' end is in closed state, and after the primer is heated through reaction to the annealing temperature, the hairpin structure is opened for the primer to begin extending. The present invention has raised detection sensitivity and specificity, high reliability of clinical judgment of tumor cell course and prognosis, and important significance in preventing infection.

Description

technical field [0001] The invention relates to a method for detecting cell division cycle protein gene 6 in tumors by using improved primers Background technique [0002] With the development of molecular biology techniques, gene diagnosis and gene therapy are more and more widely used in clinic, such as the development and application of polymerase chain reaction (polymerase chain reaction, PCR). However, due to the characteristics of PCR technology itself, false positives and false negatives are prone to occur in clinical applications. In order to avoid these errors, many specific changes, designs, analyzes and experimental verifications should be made according to the characteristics of different nucleic acid fragments in the actual application of PCR. . This patent has made improvements in primer design to detect tumor cell division cycle protein gene 6. In order to improve its specificity and prevent pollution, specific design and experimental verification have been ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张曼
Owner 张曼
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