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Primer combination for monitoring free tumor DNA, kit and application

A combination of primers and kit technology, applied in the field of medical biology, can solve the problems of unfavorable rapid detection and standardized detection, cumbersome detection process steps, high detection cost, etc., and achieve the effect of shortening the detection time, shortening the detection time, and improving the detection efficiency

Inactive Publication Date: 2018-11-27
邹畅 +3
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0013] It takes a long time for high-throughput next-generation sequencing to detect, and it generally takes one week to issue the test results. The detection process steps are cumbersome, the operation is cumbersome, and it takes a long time, which is not conducive to rapid detection and standardized detection, and the detection cost is relatively high. High, the detection cost is more than 1,000 yuan / sample

Method used

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  • Primer combination for monitoring free tumor DNA, kit and application
  • Primer combination for monitoring free tumor DNA, kit and application
  • Primer combination for monitoring free tumor DNA, kit and application

Examples

Experimental program
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Effect test

Embodiment

[0062] The method for monitoring free tumor DNA is carried out according to the following steps:

[0063] 1. Primer design:

[0064] After looking up the gene sequence of human LINE on the Genbank website, design specific amplification primers:

[0065]

[0066]

[0067] 2. Sample extraction:

[0068] First, collect 5mL of peripheral blood from tumor patients by using EDTA anticoagulant tubes, and record the sample information. If the peripheral blood samples are processed within 2 hours, they should be stored and transported at room temperature. If they are more than 2 hours After sample processing, low-temperature storage and transportation are required.

[0069] Second, the peripheral blood samples collected from tumor patients were centrifuged at 1300g, 4°C, for 20 minutes, the supernatant was drawn into a new centrifuge tube, and then centrifuged at 12000g, 4°C, for 10 minutes, to obtain peripheral blood plasma. Take 200 μL for the next step immediately, and stor...

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Abstract

The invention provides a primer combination for monitoring free tumor DNA, a kit and application. The primer combination is composed of one or two of a first primer pair and a second primer pair, thefirst primer pair is composed of an upstream primer with a base sequence shown as SEQ ID No.1 and a dowmstream primer with a base sequence shown as SEQ ID No.2, and the second primer pair is composedof an upstream primer with a base sequence shown as SEQ ID No.3 and a dowmstream primer with a base sequence shown as SEQ ID No.4. The kit comprises the primer combination. The kit has the advantagesof being quick, simple, convenient, high in specificity, sensitivity and reliability, low in detection cost, short in time, high in detection efficiency and accuracy and low false positive rate.

Description

technical field [0001] The invention relates to the field of medicine and biology, in particular to a primer combination, a kit and an application thereof for monitoring free tumor DNA. Background technique [0002] Plasma free DNA, namely cfDNA (cell-free DNA), is the free DNA in plasma, mainly derived from normal cells, abnormal cells (such as tumor cells) and external (such as viral DNA). The integrity detection of cfDNA can be used for early diagnosis, medication and monitoring of cancer. However, the current technical solutions mainly focus on the free DNA detection solution based on high-throughput sequencing, which leads to high cost and long time for this solution. [0003] The method of using high-throughput sequencing in the prior art is mainly carried out according to the following steps: [0004] 1. Take peripheral blood samples for tumor patients. [0005] 2. Plasma separation is performed on the collected patient blood samples by centrifugation to obtain pat...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6886C12Q2600/158C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 邹畅程险峰洪马林李伟
Owner 邹畅
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