DNA virus reference material and preparation method and application thereof

A DNA virus and reference material technology, applied in the field of molecular biology and animal quarantine, can solve problems such as inability to monitor quality, affect results, and inability to control reagent quality, achieving accurate copy number determination, good applicability, and a wide range of applications Effect

Inactive Publication Date: 2015-12-30
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many DNA viruses mainly use nucleic acid substances or plasmids as standards in the detection process. These standards face several important problems in the use process: (1) It is impossible to monitor the quality of the operations performed during the sample processing process
(2) Unable to perform quality control on reagents used in sample processing
Once there is a problem in these two links, it will directly affect the judgment of the result

Method used

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  • DNA virus reference material and preparation method and application thereof
  • DNA virus reference material and preparation method and application thereof
  • DNA virus reference material and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Preparation and application of embodiment 1 African swine fever virus reference material

[0038] 1. Materials

[0039] sf9 cells, pFastBac1 vector, E. coli DH10Bac cells, restriction enzymes.

[0040] 2. Primers

[0041] Primer5.0 was used to design primers according to the P72 gene sequence registered in GenBank (African swine fever virus E75, GenBank: FN557520.1), and restriction sites EcoRI and XhoI were introduced into the upstream and downstream primers respectively, and the upstream primer p72-Bac- F: 5'-CG GAATTC atggcatcaggagggagc-3 (EcoRI), downstream primer p72-Bac-R: 5'-CC CTCGAG ttaggtactgtaacgcagc-3 (XhoI). Identification of recombinant virus M13 primers: upstream primer M13For: 5'-GTTTCCCAGTCACGAC-3', downstream primer M13Rev: 5'-CAGGAAACAGCTATGAC-3'.

[0042] 3. Amplification of P72 gene and plasmid construction

[0043] In this example, the P72 positive clone plasmid was used as a template, and the primers: p72-Bac-F, p72-Bac-R were used for PCR...

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Abstract

The invention provides a DNA virus reference material and a preparation method and application thereof. The DNA virus reference material is recombinant viruses which are obtained by recombining coding genes of nucleocapsid protein in the DNA viruses into baculovirus to be expanded and purified. The DNA virus reference material prepared from baculovirus contains a virus structure and can serve as a parallel contrast in the sample processing and extracting process, and conduct quality control on the operation technology. In addition, the prepared reference material can conduct quality control on agents used in the sample processing process and provide powerful guarantees for effectiveness of experimental results.

Description

technical field [0001] The invention relates to the fields of molecular biology and animal quarantine, in particular to a DNA virus reference material and its preparation method and application. Background technique [0002] DNA virus is a kind of virus that is extremely harmful to animals. Common animal DNA viruses include African swine fever virus that infects pigs, and iridescent virus that infects fish and amphibians. [0003] Among them, African swine fever virus is a class of acute and severe infectious diseases. African swine fever (African swine fever, ASF) is an acute, febrile, highly contagious disease of pigs caused by African swine fever virus (ASFV). Can spread across borders. African swine fever virus is a DNA virus with an icosahedral structure. The virus is mainly composed of 4 layers: a central nucleolus, a nucleocapsid, an inner envelope, and an icosahedral virus nucleocapsid. In addition, the viral nucleocapsid is also wrapped with an outer envelope deri...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/866C12Q1/70C12Q1/68C12R1/93
Inventor 冯春燕
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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