A kind of human herpesvirus type 4 detection kit and detection method

A human herpes virus and detection kit technology, which is applied in the biological field, can solve the problems of nucleic acid samples containing many impurities, affecting the detection accuracy of the kit, and low extraction efficiency, and achieves the improvement of nucleic acid extraction rate, high accuracy and reduction of production. cost effect

Active Publication Date: 2019-07-09
艾吉泰康(嘉兴)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • A kind of human herpesvirus type 4 detection kit and detection method
  • A kind of human herpesvirus type 4 detection kit and detection method
  • A kind of human herpesvirus type 4 detection kit and detection method

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Embodiment 1

[0028] Embodiment 1, a kind of human herpes virus type 4 detection kit

[0029] Embodiment 1 of the present invention comprises the following components:

[0030] ①Nucleic acid release agent: add surfactant peptide to sterile water to make the final concentration reach 0.2mg / ml, add chlorhexidine acetate to make the final concentration reach 0.03mg / ml, add potassium chloride to make the final concentration reach 4.5% (w / v ) and adding hydrophilic graphene to make the final concentration reach 1.2% (w / v).

[0031] The preparation method of hydrophilic graphene: take the graphite oxide prepared by the Hummers method that has removed residual ions after dialysis, and prepare an aqueous solution of 0.5 mg / ml, according to the quality of graphite oxide: hexamethylenetetramine is 2: 1 Hexamethylenetetramine was added, stirred and refluxed at 100°C for 12 hours, and a uniformly dispersed hydrophilic graphene dispersion was obtained.

[0032] ②Nucleic acid extraction solution contai...

Embodiment 2、 Embodiment 1

[0042] Embodiment 2, the method for detecting EBV with the kit described in embodiment 1

[0043] 1. Preparation of samples to be tested

[0044] Preparation of serum samples: select freshly drawn venous blood, place it at room temperature for 25-35 minutes, centrifuge at 2000 rpm / min for 2-4 minutes, absorb the supernatant, and transfer it to a clean EP tube for later use.

[0045] 2. Nucleic acid extraction

[0046] A) Lyse the virus: Take four centrifuge tubes and add 6 μl nucleic acid release agent and 12 μl of the sample to be tested prepared in step 1), 6 μl nucleic acid release agent and 8 μl EBV negative control, 6 μl nucleic acid release agent and 8 μl EBV positive control, and 6 μl Nucleic acid release agent and 8 μl EBV quantitative reference substance, marked, mixed thoroughly, placed at room temperature for 3 minutes, set aside;

[0047] B) Magnetic bead adsorption: Add 100 μl of nucleic acid extraction solution containing magnetic beads and 4 μl of internal sta...

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Abstract

The invention belongs to the field of biotechnology, and specifically relates to a human herpesvirus 4 detecting kit and a detecting method. The human herpesvirus 4 detecting kit comprises a nucleic releaser, magnetic bead containing nucleic extracting liquid and PCR (Polymerase Chain Reaction) reacting liquid, wherein the nucleic releaser comprises surfactin, chlorhexidine acetate, 2-6% of potassium chloride and hydrophilic graphene; the magnetic bead containing nucleic extracting liquid comprises 4-hydroxyethylpiperazine ethane sulfonic acid, sodium chloride and magnetic balls; the PCR reacting liquid comprises an upstream primer, a downstream primer, and a detector; the upstream primer and the downstream primer are used for target polynucleotide amplification; the detector is used for detecting target polynucleotide. The kit has the advantages of being simple to operate, high in detection sensitivity, high in accuracy, high in specificity and wide in detection range; reliable experiment basis is provided for clinically diagnose EBV (Epstein-Barr Virus) infection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a human herpes virus type 4 detection kit and detection method. Background technique [0002] The fourth type of human herpes virus is Epstein-Barr virus (EBV). EBV is a herpes virus that is ubiquitous in the human body. Once a person is infected, he may carry it for life. EBV infection mainly causes infectious mononucleosis in children and is closely related to the occurrence of many tumors, which seriously threatens the health of human beings, especially children, so it has always been concerned. [0003] At present, the detection of Epstein-Barr virus DNA in blood mainly adopts polymerase chain reaction (PCR) technology. Fluorescent quantitative PCR, also known as real-time quantitative PCR, is a new PCR technology developed in recent years. It can monitor the fluorescently-labeled PCR reaction optically throughout the whole process, dynamically and uninterruptedly, s...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12R1/93
CPCC12Q1/6806C12Q1/686C12Q1/705C12Q2563/143C12Q2563/149C12Q2545/101C12Q2563/107
Inventor 陈立国邹伟权张亚丽李庆祥母润红王涛苏焱
Owner 艾吉泰康(嘉兴)生物科技有限公司
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