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Nasopharyngeal carcinoma related EB virus detection kit and detection method

A detection kit, Epstein-Barr virus technology, applied in the biological field, to achieve the effect of easy operation

Inactive Publication Date: 2015-11-11
慈溪市人民医院 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are several kits for detecting EBV-DNA on the market abroad. The detection method is mainly fluorescent immunohybridization (HIGH). The ELISA method is mainly used for domestic detection of EBV. There is no kit approved for the detection of EBV-DNA on the market.

Method used

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  • Nasopharyngeal carcinoma related EB virus detection kit and detection method
  • Nasopharyngeal carcinoma related EB virus detection kit and detection method
  • Nasopharyngeal carcinoma related EB virus detection kit and detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0062] Embodiment 1 A detection method for detecting nasopharyngeal carcinoma-associated Epstein-Barr virus using the above kit: comprising the following steps:

[0063] (1) Paraffin specimen extraction:

[0064] A. Dewaxing:

[0065] ①. Add 1ml xylene and vortex vigorously for 10s;

[0066] ②, room temperature centrifugation, 12000rpm, 2min;

[0067] ③. Carefully absorb the supernatant and discard it. Be careful not to remove the tissue. If there are too many paraffin impurities, repeat it once;

[0068] ④. Add 1ml of absolute ethanol and shake vigorously;

[0069] ⑤. Centrifuge at room temperature, 12000rpm, 2min;

[0070] ⑥ . 1. Carefully absorb the supernatant, discard it, and be careful not to remove the tissue;

[0071] ⑦ . , Put it in a metal bath, and bake at 37°C until completely dry.

[0072] B. Extraction:

[0073] ①. Add 180 μL BufferATL, 20 μl proteinase K (proteinase K), shake vigorously and mix well;

[0074] ②. Place at 56°C for 1 hour, or until the t...

Embodiment 2

[0095] Embodiment 2 A detection method utilizing the above kit to detect nasopharyngeal carcinoma-associated Epstein-Barr virus: comprising the following steps:

[0096] (1) Extraction of tissue samples

[0097] Experimental Materials:

[0098] Qiagen Paraffin Extraction Kit (Product No. 51304), absolute ethanol, metal bath, high-speed centrifuge, micropipette;

[0099] Experimental steps:

[0100] ①. Add 20 μl proteinase K (proteinase K), add an appropriate amount of surgical tissue, and grind until pulverized with a tissue grinder;

[0101] ② Add 200 μL of BufferAL, shake vigorously for 15 seconds, place at 56°C for 10 minutes, and centrifuge instantly;

[0102] ③. Add 200 μl absolute ethanol, shake and mix again for 15 seconds, and then centrifuge instantly;

[0103] ④. Carefully transfer the liquid to the QIAamp Minispin column (there is a 2ml EP tube under the spin column to collect the waste liquid), and centrifuge at 8000rpm for 1min. Discard the liquid in the wast...

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Abstract

The invention relates to a nasopharyngeal carcinoma related EB virus detection kit. The detection kit comprises (1) an EB-PCR reaction solution which comprises buffer, dNTPs and taq enzyme, (2) a first EB primer probe solution, (3) a second EB primer probe solution, (4) a critical positive quality control material, namely recombinant plasmids containing amplified target gene fragments, (5) a positive quality control material, namely recombinant plasmids containing amplified target gene fragments, (6) a negative quality control material, namely DEPC treated water. According to the nasopharyngeal carcinoma related EB virus detection kit provided by the invention, a nucleic acid amplification technology and a Taqman fluorescent probe technique are combined, so that whether EB viral nucleic acid exists or not can be judged by detecting amplification products, and the viral load can be detected through a quantitative standard curve; besides, the risk of incidence of nasopharyngeal carcinoma can be judged according to viral load, and the detection kit has the advantages of being peculiar, sensitive, rapid and convenient to operate.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a nasopharyngeal carcinoma-related Epstein-Barr virus detection kit and a detection method. Background technique [0002] Epstein-Barr virus (EBV) infection is relatively common all over the world, and more than 90% of adults have been infected. Epstein-Barr virus infection can cause infectious mononucleosis, which is closely related to Burkitt's lymphoma, Hodgkin's disease and nasopharyngeal carcinoma (NPC). Current studies have found that EBV infection is associated with a variety of human tumors. Including Burkitt's lymphoma, Hodgkin's lymphoma, gastric cancer, nasopharyngeal cancer, etc. EBV plays a very important role in the occurrence of these tumors, so in 1997, EBV was classified as the first type of carcinogen. In recent years, researchers have found that plasma free EBV-DNA is a reliable monitoring indicator, which plays a very good role in the diagnosis, monitoring of c...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/701C12Q1/6844C12Q1/6886C12Q1/70C12Q2561/101C12Q2561/113
Inventor 龚梁张贻琛
Owner 慈溪市人民医院