Probe used for EBV detection and detection method

A probe and reaction technology, applied in the field of genetic engineering, can solve the problems of low extraction efficiency, affecting the detection accuracy of kits, and nucleic acid samples containing many impurities, and achieves the effects of high coverage depth, low cost, and fast data analysis.

Inactive Publication Date: 2020-04-28
HANGZHOU LC BIOTECH
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Problems solved by technology

Although nucleic acid extraction with a nucleic acid release agent has the advantages of quickness and simplicity, the nucleic acid sample contains many impurities, and the overall extraction efficiency is not high, which affects the detection accuracy of the kit.

Method used

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  • Probe used for EBV detection and detection method
  • Probe used for EBV detection and detection method
  • Probe used for EBV detection and detection method

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Experimental program
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Embodiment approach

[0052] General implementation plan: a probe and detection method for EBV detection, comprising the following steps:

[0053] (1) Prepare the probe;

[0054](2) Prepare the DNA of cervical exfoliated cell samples with complete electrophoresis main band and no obvious degradation;

[0055] (3) Fragmentation of genomic DNA using ultrasonic physical fragmentation method;

[0056] (4) Purifying the fragmented product in step (3) with LC magnetic beads;

[0057] (5) constructing the fragmented DNA into an Illumina genome library;

[0058] (6) Amplify the genomic library by PCR reaction and use LC magnetic beads to purify the amplified product;

[0059] (7) Hybridizing the probe prepared in step (1) with the genomic library in step (6);

[0060] (8) capture the hybridization product with streptavidin magnetic beads and wash the non-specific binding;

[0061] (9) PCR amplification of target DNA fragments bound to the streptavidin magnetic beads;

[0062] (10) High-throughput seq...

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Abstract

The invention discloses a probe used for EBV detection and a detection method. Two rounds of PCR reaction are needed in the probe preparation process. The detection method based on an NGS technology comprises the following steps: preparing a DNA template, fragmenting genome DNA, purifying a fragmented product, constructing an introduction group library, carrying out PCR amplification on the genomelibrary, hybridizing a probe with the genome library, allowing magnetic beads to capture a hybridization product, carrying out high-throughput sequencing on an obtained captured product, and analyzing the EBV infection condition in a to-be-detected sample. The probe and the method disclosed by the invention have the advantages of low cost, fast data analysis, high coverage depth, high detection accuracy and the like, are applicable to early screening and postoperative diagnosis of diseases caused by EBV, and provide a guidance for personalized medication.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, and more specifically relates to a probe and a detection method for EBV detection. Background technique [0002] Human herpesvirus type IV (Epstein-Barr virus, EBV), also known as Epstein-Barr virus, is one of the most common viruses that can cause human diseases. It is a double-stranded linear DNA with a genome length of about 172kb. Its shape is similar to other herpes viruses. Viruses are similar, round, 180nm in diameter, and the basic structure includes three parts: nucleus, capsid and envelope. It can lead to host chromosomal translocation and rearrangement, affect important signaling pathways such as cell proliferation, migration, and apoptosis through LMP1 and miRNA, and achieve immune escape through LMP2. EBV is considered to be related to many diseases such as invasive breast cancer, gastric cancer, nasopharyngeal cancer, colorectal cancer, malignant blood tumors, etc. In A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6874C12N15/11
CPCC12Q1/705C12Q1/6874C12Q2531/113C12Q2535/122C12Q2563/143C12Q2563/149
Inventor 潘石玄伟钱刚蔡霖霖李振郎秋蕾
Owner HANGZHOU LC BIOTECH
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