Kit and preparation method for detecting foot-and-mouth disease type a, type o and asia 1 virus

A detection kit and a technology for foot-and-mouth disease virus, which are applied to primers of O-type and Asia1-type viruses, detect the field of foot-and-mouth disease type A, and can solve problems such as inability to distinguish between serotypes

Active Publication Date: 2016-03-30
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 Asia1 in the cell culture supernatant , but cannot distinguish between these serotypes

Method used

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  • Kit and preparation method for detecting foot-and-mouth disease type a, type o and asia 1 virus
  • Kit and preparation method for detecting foot-and-mouth disease type a, type o and asia 1 virus
  • Kit and preparation method for detecting foot-and-mouth disease type a, type o and asia 1 virus

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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-Asia1 published by NCBI, select conserved regions to design specific primers, and form specific chimeric primers by adding universal primers. The universal primer sequence belongs to A non-biologically derived nucleotide sequence, in addition to synthesizing a universal primer sequence, and adding a Cy5 fluorescent label 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...

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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 type A, type O and Asia1 virus, a kit containing 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). Huge political and economic losses, so the World Health Organization (OIE) listed it as the first class A infectious disease. FMDV can be divided into seven serotypes, namely A, O, C, Asia1, SAT1, SAT2 and SAT3, each of which has many different subtypes, and there is no crossover between the serotypes 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 control of the spread of the disease, the differential diagnosis of foot-and-mouth disease virus has always been a research hotspot...

Claims

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Application Information

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Patent Type & Authority Patents(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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