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Method for detecting Ct values of PCA3 and PSA in urine

A urine and real-time detection technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of low tumor specificity, achieve excellent sensitivity and specificity, and rigorous PCR amplification procedures Effect

Pending Publication Date: 2019-06-25
HANGZHOU YORK BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the low tumor specificity of PSA, scholars have been looking for new prostate cancer-specific tumor markers

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The method for the Ct value of PCA3 and PSA in the detection urine of the present embodiment comprises the following steps:

[0034] Step 1, design primers and probes of PCA3 and PSA, including:

[0035] PCA3 primers: 01F: GACTACAGACATCTCAATG; 02R: CCAGGGATCTCTGTGCT;

[0036] PCA3 Taqman probe 03Q:FAM-AACATCACTAGAAACAGCAAG-TAMRA;

[0037] PSA primers: 04F: CGATATGAGCCTCCTGAA; 05R: TGGTCCCCAGTGCTGG;

[0038] PSA Taqman probe 06Q: FAM-ACGGATGCTGTGAAGGTCATGGACCT-TAMRA.

[0039] Step 2. Extract the RNA in the urine, specifically: after adding chloroform to separate the two phases, precipitate the RNA with isopropanol, wash it several times with ethanol, dry it, add water to dissolve it, pack it separately, and store it in a -80°C refrigerator live.

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Abstract

The invention discloses a method for detecting Ct values of PCA3 and PSA in urine. The method comprises the first step of designing primers and probes of the PCA3 and the PSA; the second step of extracting RNA in the urine; the third step of designing a PCR amplification system, then conducting PCR amplification and detecting the PCR process in real time by fluorescence signals during the PCR amplification process; the fourth step of recording the Ct values of the PCA3 and the PSA. The Ct values of the PCA3 and the PSA can respectively express the expression levels of PCA3 mRNA and PSA mRNA, and the ratio of PCA3 RNA to PSA RNA (PCA3 mRNA) / (PSA mRNA)*1000 can be accurately quantified according to the PCA3 score. The expression of the PCA3 RNA can be directly quantified to directly guide whether prostate puncture is needed.

Description

technical field [0001] The invention relates to a method for detecting the Ct value of PCA3 and PSA in urine. Background technique [0002] According to my country's "Prostate Cancer Diagnosis and Treatment Standards (2018 Edition)", the incidence of prostate cancer ranks second among male malignancies worldwide, and the incidence of prostate cancer in the United States has surpassed lung cancer, becoming the number one hazard to men's health. tumor. In the United States, the incidence peak begins at the age of 50, and nearly 91% of new prostate cancer cases are clinically localized prostate cancer, and the 5-year survival rate after receiving standard treatment is close to 100%. In my country, the peak incidence of prostate cancer begins at the age of 60 (in cities), and 70% of new cases are patients with locally advanced or extensive metastases. These patients cannot receive local radical treatment, and their prognosis is poor. Therefore, at this stage in our country, it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
Inventor 林忠旺赵培雅
Owner HANGZHOU YORK BIOTECH CO LTD