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Method for preparing PCA3 (prostate cancer antigen 3) quantitative detection standard

A quantitative detection and standard technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as low specificity and negative puncture, and achieve the effect of high sensitivity and specificity

Pending Publication Date: 2019-06-21
HANGZHOU YORK BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the traditional prostate biopsy has high sensitivity, it has low specificity and is prone to unnecessary negative punctures. Therefore, it is necessary to find a method that can effectively diagnose prostate cancer

Method used

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  • Method for preparing PCA3 (prostate cancer antigen 3) quantitative detection standard
  • Method for preparing PCA3 (prostate cancer antigen 3) quantitative detection standard
  • Method for preparing PCA3 (prostate cancer antigen 3) quantitative detection standard

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Embodiment 1

[0026] The method for preparing PCA3 quantitative detection standard substance of the present embodiment, comprises the following steps:

[0027] Step 1: Synthesizing PCA3 DNA, and the synthesized sequence is coded as PCA3-1;

[0028] Step 2: PCR amplification; define the upstream and downstream primer codes as PCA3-F and PCA3-R respectively, and the 25ul reaction system is:

[0029]

[0030] The PCR amplification program specifically includes the following steps:

[0031] S1, after configuring the components required for the PCR reaction, preheat the PCR instrument at 95°C for 5 minutes to fully denature the template DNA, and then enter the amplification cycle;

[0032] S2, Amplification cycle: In each cycle, first keep at 95°C for 10s to denature the template, then lower the temperature to 55°C for 10s to fully anneal the primers to the template, and finally keep at 72°C for 30s to make the primers on the template Up extension, DNA synthesis, complete a cycle, repeat th...

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Abstract

The invention discloses a method for preparing a PCA3 (prostate cancer antigen 3) quantitative detection standard. The method includes the following steps: step 1, synthesizing PCA3 DNA; step 2, performing PCR amplification; step 3, purifying a PCR product; step 4, performing in-vitro transcription to obtain RNA; step 5, purifying a transcribed product; step 6, quantifying the RNA standard. The PCA3 quantitative detection standard prepared by the method can be used as a positive quality control or a standard curve, the gene copy number is quantified by a real-time fluorescence quantitative method, and the sensitivity and specificity are high.

Description

technical field [0001] The invention relates to a method for preparing a PCA3 quantitative detection standard product. Background technique [0002] At this stage in our country, the main content of prostate cancer screening is as follows: It is recommended that men over 50 years old, or men over 45 years old with a family history of prostate cancer, be tested with PSA (PSA test) on the premise of fully informing the screening risks Based prostate cancer screening, in order to reduce mortality by early detection. PSA is widely used clinically as a marker of prostate cancer. Although it can effectively diagnose some prostate cancer patients, benign prostatic hyperplasia, prostatitis, prostate massage, and transrectal ultrasonography can all lead to an abnormal increase in PSA levels, and there are many studies It shows that when the PSA is in the gray area, that is, 4-10ng / ml, the diagnostic rate is only 25%. Therefore, the tumor specificity of PSA is not high, and scholars...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806
Inventor 林忠旺赵培雅
Owner HANGZHOU YORK BIOTECH CO LTD
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