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Assay method for EBV (Epstein-Barr virus)

A detection method, virus technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganism measurement/inspection, etc., can solve the problems of incompleteness and low sequencing throughput, and achieve low false negatives and good repeatability , the effect of high accuracy

Pending Publication Date: 2018-02-23
JIAXING YUNYING MEDICAL INSPECTION CO LTD
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AI Technical Summary

Problems solved by technology

In the field of molecular diagnosis, the most direct and clear technology is gene sequencing. The traditional gene sequencing method is carried out by the Sanger termination method. The disadvantage is that only a single gene sequence can be read at a time, and the sequencing throughput is too low and not comprehensive.

Method used

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  • Assay method for EBV (Epstein-Barr virus)
  • Assay method for EBV (Epstein-Barr virus)

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0025] The invention provides a kind of Epstein-Barr virus (EBV) integrated identification high-throughput sequencing detection method, comprises the steps:

[0026] 1. Sample library preparation:

[0027] (1) Ultrasound fragmentation: the initial amount is 3 μg, add RNA-free-water to 100 μl and dilute to 30 ng / μl. SCIENTZ08-Ⅲ cup-type ultrasonic cell pulverizer was used for ultrasonic fragmentation, and the setting parameters were: power 70%, interruption for 3s, stop for 1s, and cycle for 30-60min.

[0028] (2) Take out the magnetic beads half an hour in advance to return to room temperature, shake and mix, take 180ul of magnetic beads and add them to the PCR product interrupted by ultrasound, beat and mix, and incubate in the greenhouse for 5 minutes.

[0029] (3) Put the PCR single tube on the magnetic stand, let it stand for 5 minutes, remove the supernatant, keep the PCR tube on the magnetic stand, add 200ul of 80% ethanol (newly prepared) to rinse, let stand for 30s, r...

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Abstract

The invention discloses a high-throughput sequencing assay method for Epstein-Barr virus (EBV) integration assay. The method utilizes the genome information of the Epstein-Barr virus (EBV) in combination with the second-generation high-throughput sequencing technology to more comprehensively assay the condition of EBV infection of a patient, overcoming the defects of conventional assay methods, such as low accuracy, high false positive and poor repetitiveness. The adoption of gene sequencing can help increasing accuracy, and compared with the classical Sanger sequencing method mostly adopted at present, the assay method has the advantages of higher assay throughput, higher accuracy, lower cost, richer information and the like. With the help of the second-generation high-throughput sequencing technology, the assay method can carry out high-precision assay on the human infection of the Epstein-Barr virus, so that specific treatment can be carried out at the infection stage to reduce theharm of the disease on the human body, and thereby the occurrence of tumor is decreased.

Description

1. Technical field [0001] The invention relates to a virus detection method, in particular to a high-throughput sequencing detection method for integrated identification of Epstein-Barr virus (EBV). 2. Technical background [0002] Nasopharyngeal carcinoma (NPC) is a malignant tumor that occurs in the nasopharynx between the nasal cavity and the pharynx. It originates in the epithelium of the nasopharyngeal mucosa. Characterized by readily invasive growth and early metastasis. In my country, nasopharyngeal carcinoma is one of the seven malignant tumors with the highest mortality rate, and the average survival period from the discovery of symptoms to death is only 18.7 months. According to the classification of the World Health Organization, nasopharyngeal carcinoma is divided into keratinizing squamous cell carcinoma (95%) and nonkeratinizing carcinoma. It is currently known that the occurrence of nasopharyngeal carcinoma is related to many factors, among which Epstein-Bar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6869C12R1/93
CPCC12Q1/6869C12Q2565/519C12Q2535/122
Inventor 张道允巩子英叶建伟王伟
Owner JIAXING YUNYING MEDICAL INSPECTION CO LTD
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