A kind of sequencing primer, detection method and kit for dengue virus typing

A dengue virus and kit technology, applied in the field of biomedicine, can solve problems such as the inability to determine the virus type

Active Publication Date: 2022-05-13
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Description
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AI Technical Summary

Problems solved by technology

Even if the dengue virus is monitored by the reaction pool method, false negatives still exist, and the virus type cannot be determined

Method used

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  • A kind of sequencing primer, detection method and kit for dengue virus typing
  • A kind of sequencing primer, detection method and kit for dengue virus typing
  • A kind of sequencing primer, detection method and kit for dengue virus typing

Examples

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Comparison scheme
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Embodiment 1

[0109] Embodiment 1, detection method of the present invention

[0110] The operation process of the multiplex PCR_MS mass spectrometry scanning method of the present invention is as follows figure 1 ,details as follows:

[0111] (1) Establishment of dengue virus gene database: 4,502 complete genome sequences of dengue virus types I-IV were obtained from the NCBI website, including: 1,874 for type I, 1,478 for type II, 944 for type III and 206 for type IV. See Table 1 for details.

[0112] Table 1. Sequence source of primers designed for dengue virus DENV in PCR-MS

[0113] Table 1. Reference strains used in Multiplex PCR-MS design

[0114]

[0115]

[0116] (2) Establishment of SNP typing of dengue virus: After clustering analysis with Sequencher5.3 software, a type-specific SNP site (type I / C, type II / A, type III / T, type IV / A) design primers and probes, and cooperate with auxiliary identification. Establish dengue virus SNP typing table, see Table 2 for details.

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Abstract

The invention belongs to the field of biomedicine, and relates to a sequencing primer, a detection method and a kit for dengue virus typing. The present invention establishes a multiplex PCR-MS mass spectrometry detection method that comprehensively covers four serotypes of dengue virus and its mutant strains, and is used for laboratory diagnosis, precise genotyping and sample tracing of dengue virus.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a sequencing primer, a detection method and a kit for dengue virus typing. Background technique [0002] Dengue virus (DV) belongs to the genus Flaviviridae of the family Flaviviridae. Dengue virus particles are spherical, with a diameter of 45-55 nm. They are enveloped single-stranded positive-sense RNA viruses. The genome is about 11 kb long. The coding sequence of the entire gene is: 5' I-type cap structure, non-coding region, AUC protein ( Nucleocapsid protein) gene, M protein (membrane protein) gene, E protein (envelope protein) gene, NS1-2a-2b-3-4a-4b-5 gene, non-coding region 3'. There are four serotypes of dengue virus (DENV-1, DENV-2, DENV-3 and DENV-4), all of which can infect humans and cause recessive infection, dengue fever (dengue fever, DF) and dengue hemorrhage Fever (denguehemorrhagicfever, DHF), severe can also lead to dengue shock syndrome (dengueshocksyndrome, DSS),...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6872C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6872C12Q2600/16C12Q2600/156C12Q2531/113C12Q2537/143C12Q2521/525C12Q2533/101C12Q2565/627
Inventor 曹晓梅林健李颖常宇桐张丽萍刘莹莹牛羽李慧鹏
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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