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Biomarker combinations, kits and applications for early screening of prostate cancer

A biomarker, prostate cancer technology, applied in the field of biomarker combination for early screening of prostate cancer, can solve the problems of poor specificity, increased pain and injury, low sensitivity of prostate cancer detection, etc., and achieves good stability and specificity. Strong and efficient effect

Active Publication Date: 2022-02-22
求臻医学科技(浙江)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the shortcomings of the above-mentioned background technology, the present invention provides a biomarker combination, kit and application for early screening of prostate cancer, which solves the problem of low sensitivity and poor specificity of prostate cancer detection, and avoids excessive diagnosis and treatment that increase the risk of Necessary Pain and Injury Issues

Method used

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  • Biomarker combinations, kits and applications for early screening of prostate cancer
  • Biomarker combinations, kits and applications for early screening of prostate cancer
  • Biomarker combinations, kits and applications for early screening of prostate cancer

Examples

Experimental program
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Effect test

Embodiment 1

[0050] Example 1 Kit for Early Screening of Prostate Cancer

[0051] Including Qiagen urine free DNA extraction kit, HinP1I, HpaII, AciI, HpyCH4IV methylation-sensitive endonuclease system, 5 pairs of target-specific primers and internal reference primers, 5 pairs of specific primer probes and internal reference primer probes;

[0052] Wherein the sequences of the 5 pairs of target-specific primers are as follows:

[0053] Seq ID NO.1F ACCGAGAAGAAAAGGCCGTACTC,

[0054] Seq ID NO. 1R TGTGAACCTTTCGCGGCTCGAC;

[0055] Seq ID NO. 2F AGTTGCTGACGCAGTTCCTTC,

[0056] Seq ID NO.2R CCACGCCAGACTGTAGAGCAC;

[0057] Seq ID NO. 3F GGAATTCTTTTGCTGCGCTG,

[0058] Seq ID NO. 3R CAAGGCCCGGCACTTTTCA;

[0059] Seq ID NO. 4F CCGAAGCCCACGTAGCCTG,

[0060]Seq ID NO.4R CACCGGACCCTCCTGGACTC;

[0061] Seq ID NO.5F CAAAGCCGCCGCTGCCAAAG,

[0062] Seq ID NO.5R CTCGGGGTCCAATAGTAGCGGGTAC;

[0063] The sequences of internal reference primers are as follows:

[0064] Seq ID NO.6F CCTACGAAAACCTCACGG...

Embodiment 2

[0074] Example 2 Method for detecting changes in ctDNA methylation level

[0075] 1. Take 10 mL of urine and use Qiagen Urine Free DNA Extraction Kit (Cat: 954154) to extract free cfDNA;

[0076] 2. Take 20 ng cfDNA and 20 μL HinP1I, HpaII, AciI, HpyCH4IV four methylation-sensitive endonuclease systems (final concentration 10 U / μL), incubate at 37°C for 16 h, and inactivate the enzyme at 80°C for 20 min;

[0077] 3. Use all the incubated products as templates, 5 pairs of target-specific primers (50 nM each) plus 1 internal reference primer (10 nM) to configure the system, set the following table 1 to set the program and perform the amplification program in a common PCR instrument ;

[0078] Table 1

[0079]

[0080] 4. Take 1 μL of the amplification product from the previous step as a template, add 5 pairs of target-specific primers (0.25 μM each), 5 specific primer probes (0.1 μM each), plus 1 internal reference primer (0.05 μM) and internal reference Primer probe (0.02...

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Abstract

The present invention relates to the field of molecular biology technology, in particular to a biomarker combination, kit and application for early screening of prostate cancer. Site-related 5 pairs of target-specific primers and corresponding internal reference primers, specific primer probes. Compared with the prior art, the present invention utilizes the property that methylation-sensitive endonucleases cannot cut methylation sites, degrades non-tumor-derived cfDNA, and specifically amplifies tumor-derived ctDNA, greatly improving detection sensitivity, and can be used for early invasiveness Early screening, early diagnosis, efficacy evaluation and follow-up, and prognosis assessment of low-grade, asymptomatic occult prostate cancer.

Description

technical field [0001] The invention relates to the field of molecular biology technology, in particular to a biomarker combination, kit and application for early screening of prostate cancer. Background technique [0002] Prostate cancer is the malignant tumor with the highest incidence rate in European and American men, and it ranks second in the male tumor mortality rate in the United States, second only to lung cancer. According to statistics, there were 241,740 newly diagnosed prostate cancer patients in the United States in 2012, accounting for 29% of all new tumor patients; 28,170 people died of prostate cancer, accounting for 9% of all tumor deaths. In recent years, the incidence of prostate cancer in my country has also been increasing. [0003] The only marker clinically used for the diagnosis of prostate cancer and the monitoring of recurrence after treatment is the serological marker prostate-specific antigen (PSA), but about 1 / 3 of the patients detected by PSA-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/154C12Q2600/166C12Q2600/16C12Q2600/118C12Q2531/113C12Q2521/331C12Q2537/143C12Q2545/101
Inventor 宋莹莹张怡然王昊昊张腾龙段小红顾丽清
Owner 求臻医学科技(浙江)有限公司
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