Reagent box for detecting aftosa A type, O type and Asia 1 type viruses and preparing method thereof

A detection kit and technology of foot-and-mouth disease virus, which is applied to the primers of type O and Asia 1 virus, and the field of detection of type A of foot-and-mouth disease, which can solve the problem of inability to distinguish serotypes

Active Publication Date: 2014-12-10
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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AI Technical Summary

Problems solved by technology

The sensitivity of the test strip to detect the FMDV antigen in the epithelial suspension test is the same as that of the conventional antigen ELISA, and has 100% equivalent sensitivity to the FMDV serotypes O, A, C and Asia 1 in the cell culture supernatant sex, but cannot distinguish between these serotypes

Method used

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  • Reagent box for detecting aftosa A type, O type and Asia 1 type viruses and preparing method thereof
  • Reagent box for detecting aftosa A type, O type and Asia 1 type viruses and preparing method thereof
  • Reagent box for detecting aftosa A type, O type and Asia 1 type viruses and preparing method thereof

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Embodiment Construction

[0038] The present invention will be explained in detail below in conjunction with the examples.

[0039] 1. Preparation of Sequences

[0040] According to the GeXP primer design requirements and the genomes of FMDV-A, FMDV-O, and FMDV-Asia 1 published by NCBI, the conserved regions were selected to design specific primers, and specific chimeric primers were formed by adding universal primers. It is a non-biologically derived nucleotide sequence. In addition, a universal primer sequence is synthesized, and a Cy5 fluorescent label is added to the 5' end of the upstream universal primer.

[0041] 2. Viral Genome Extraction

[0042] After the monolayer cells grow to more than 80% confluent, inoculate the virus, incubate at 37°C for 30 minutes, and discard the venom. Add cell maintenance solution, 37°C 5% CO 2 Cultivate, observe cytopathic changes (CPE) regularly, and collect the poison after more than 90% of the cells appear CPE. The virus was repeatedly frozen and thawed th...

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Abstract

The invention discloses a primer capable of being used for detecting aftosa A type, O type and Asia 1 type viruses, a reagent box comprising the primer and a preparing method thereof. Six gene sequences are available in the primer for detecting the aftosa A type, O type and Asia 1 type viruses, and two universal primers are additionally arranged. Related experiments show that the nucleic acids of the aftosa A type, O type and Asia 1 type viruses can be specifically and rapidly amplified by the primer sequences, and a specific strip is detected by nucleic acid electrophoresis, but the nucleic acids of vesicular stomatitis viruses and swine vesicular disease viruses cannot be amplified under the same conditions. The primer can be used for rapidly identifying the aftosa A type, O type and Asia 1 type viruses and applied in the epidemiological research of aftosa viruses.

Description

technical field [0001] The invention relates to a primer which can be used for detecting foot-and-mouth disease A, O and Asia 1 viruses, a kit including the primer and a preparation method. Background technique [0002] Foot-and-mouth disease (FMD) is an acute and severe infectious disease of animals caused by foot-and-mouth disease virus (FMDV), which mainly harms pigs, cattle, and sheeppox cloven-hoofed animals, with a very high incidence , has caused huge political and economic losses, so it is listed as the first class A infectious disease by the World Health Organization (OIE). FMDV can be divided into 7 serotypes, namely A, O, C, Asia 1, SAT 1, SAT 2 and SAT 3, and each serotype has many different subtypes, and each serotype There is no cross-immunity between them, and there is only partial cross-immunity between subtypes of the same serotype. [0003] Since the results of rapid differential diagnosis are beneficial to the selection of specific vaccines and the contr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/701C12Q2600/16
Inventor 张强卢昌赵志荀吴国华颜新敏李应国岳华周晓黎李健朱海霞代雪玲田波芦晓立高顺平王曼
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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