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Multi-target detecting method for early-stage bladder cancer

A detection method and a bladder cancer technology are applied in the field of multi-target detection of early bladder cancer to achieve the effects of improving sensitivity and specificity, short detection time and convenient sampling

Active Publication Date: 2014-11-12
亚能生物技术(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no patent technology related to tumors by detecting methylation biomarkers in China

Method used

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  • Multi-target detecting method for early-stage bladder cancer
  • Multi-target detecting method for early-stage bladder cancer
  • Multi-target detecting method for early-stage bladder cancer

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0039] Example 1: Multi-target detection method for early cancer:

[0040] 1. Sample source:

[0041] The urine DNA samples of 4 urinary tract inflammation urine samples from 30 bladder cancer patients and 30 normal individuals were tested. The samples of bladder cancer patients came from the Urology Department of Shenzhen Second People's Hospital, and the samples of normal people came from the urine samples of volunteers. The sample information is shown in Table 1.

[0042] Table 1 Sample information of 64 cases

[0043]

[0044]

[0045]

[0046] 2. DNA extraction and detection:

[0047] 1. DNA extraction:

[0048] After obtaining the urine sample, add the urine preservation solution according to the volume ratio urine:preservation solution = 4:1, mix well and quickly freeze it at -20°C or -80°C. When extraction is required, warm the bath until the urine is clear, centrifuge at 800g for 15 minutes in a low-temperature centrifuge at 4°C, remove the supernatant,...

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Abstract

The invention relates to a multi-target detecting method for the early-stage bladder cancer, and belongs to the technical field of biological detection. The multi-target detecting method comprises the following steps of: (1) extracting DNA contained in urea of a patient; (2) carrying out hydrosulphite conversion treatment onto the obtained DNA; (3) detecting seven marker genes including EOMES, GDF15, NID2, PCDH17, POU4F2, TCF21 and ZNF154 by virtue of methylation specific fluorescence quantitative PCR, wherein ALU-C4 is used as a reference gene; (4), judging whether the patient suffers from the early-stage bladder cancer or not according to a CT value in the step (3). The multi-target detecting method disclosed by the invention has the following advantages: a plurality of biomarker genes are adopted for joint detection, and a mode of combining a plurality of markers is applied to improve detecting sensitivity and specificity on the premise of higher specificity of each marker; specific primers are designed according to the plurality of biomarker genes.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a multi-target detection method for early bladder cancer. Background technique [0002] Bladder cancer is the fifth most common cancer in the world. In most cases, about 75% of patients are in Ta or T1 stage, which is non-muscle invasive bladder cancer (NMIBC), while the remaining 25% of tumors are in muscle invasive stage T2-T4 cancer (muscle invasive bladder cancer, MIBC). About 60%-70% of patients with NMIBC tumors relapse within 3 years after resection [1-3] . Most of the relapsed patients will relapse periodically without further deterioration of the cancer grade, but about 25% of the patients will develop MIBC. Due to the high recurrence rate and risk of progression, early diagnosis, early treatment, and frequent long-term monitoring are very important to improve the survival rate of bladder cancer patients. [0003] The traditional bladder cancer examinations ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6886C12Q2600/154C12Q2523/125C12Q2531/113C12Q2545/101
Inventor 蔡志明吴松于源叶睿李巧玲王永强
Owner 亚能生物技术(深圳)有限公司
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