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Method for quantitative detection of EB virus DNA

A quantitative detection method, EB virus technology, applied in the direction of microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of unclear standards, inaccurate analysis of test results, high cost of detection methods, etc., to achieve convenient operation and analysis Accurate results and clear standards

Inactive Publication Date: 2018-11-06
CHANGSHA KINGMED MEDICAL DIAGNOSTICS INST
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Problems solved by technology

Real-time fluorescent PCR is used to detect EBV-DNA content, which solves the technical problems of existing detection methods such as high cost, poor practical operation effect, unclear standards, and inaccurate analysis of detection results.

Method used

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  • Method for quantitative detection of EB virus DNA
  • Method for quantitative detection of EB virus DNA
  • Method for quantitative detection of EB virus DNA

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Experimental program
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specific Embodiment

[0034] 1. Preparation of reagents and equipment

[0035] 1. Sample processing reagent: red blood cell lysate

[0036] EBV nucleic acid release agent 2.5ml×1 tube

[0037] Concentrate 5ml×1 tube

[0038] 2. EBV-PCR reaction solution: 912μl×2 tubes

[0039]

[0040]

[0041] The equipment to be used is:

[0042] American Applied Biology (ABI) 7300 and 7500 fluorescent quantitative PCR instruments;

[0043] Xinbeixi secondary biological safety cabinet;

[0044] Eppendorf refrigerated high-speed centrifuge;

[0045] Eppendorf sampling gun.

[0046] 2. Reagent preparation (reagent preparation area)

[0047] 1. Take out the PCR reaction solution, enzyme mixture, and internal standard from the kit, and after melting at room temperature, centrifuge at 2000rpm for a few seconds;

[0048] 2. Set the required number of tubes as n (number of tubes n = number of samples + 1 tube of negative control + 1 tube of blank control + 1 tube of high-value quality control + 1 tube of l...

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Abstract

The invention discloses a method for quantitative detection of EB virus DNA and relates to the technical field of biological detection. The method for quantitative detection of EB virus DNA comprisesthe steps that a nucleic acid releasing agent is adopted for rapid splitting decomposition, EBV-DNA in a sample is released, different extracting modes are adopted for plasma samples, peripheral bloodsamples, pharyngeal swab samples and negative positive controls and are assisted by components such as PCR reaction liquid, a real-time fluorescent quantitative PCR detection technology is applied ona fluorescent quantitative PCR instrument, and quantitative detection of EB virus DNA is achieved through change of fluorescence signals. An EB virus can be determined qualitatively and quantitatively according to accurate reported values of EBV DNA. The method has the advantages of being rapid, accurate and convenient to operate and the like.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and specifically refers to a quantitative detection method of Epstein-Barr virus DNA. Background technique [0002] Epstein-Barr virus (EBV), also known as human herpesvirus type 4 (HHV-4), was first successfully established by epstein and barr in 1964 by in vitro suspension culture of Burkitt African children's lymphoma cells, and in the cell smear of the established strain Herpes virus particles were observed by electron microscopy. When EB virus infects the human body, it first proliferates in the oropharyngeal epithelial cells, and then infects B lymphocytes. The latent Epstein-Barr virus is activated to form recurrent infection. Therefore, detecting the content of Epstein-Barr virus is of great significance for disease screening and auxiliary diagnosis. [0003] In recent years, the relevant research on EB virus detection methods is mainly ELISA detection method and EB virus ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686
CPCC12Q1/686C12Q1/705C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 刘岱璿唐笑
Owner CHANGSHA KINGMED MEDICAL DIAGNOSTICS INST
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