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Multiple RT-PCR identification and detection reagent for animal vesicular disease and preparation method and use thereof

A RT-PCR, identification and detection technology, applied in the field of biological preparations, can solve the problems of not establishing a stable method to identify three viruses at the same time, and taking a long time to achieve safe use, a wide range of sample applications, and no biological safety. hidden effect

Inactive Publication Date: 2008-01-02
CHECKOUT & QUARANTINE TECH CENT YUNNAN ENTRY &EXIT CHECKOUT & QUARANTINE BUR
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Problems solved by technology

The clinical symptoms of the three types of vesicular diseases are indistinguishable, and differential diagnosis must be passed. At present, the isolation of pathogens and conventional serological methods are mostly used for the diagnosis of the three types of diseases, which takes a long time, and some of them use PCR technology for separate identification. However, a stable method for simultaneous identification of the three viruses has not been established

Method used

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  • Multiple RT-PCR identification and detection reagent for animal vesicular disease and preparation method and use thereof
  • Multiple RT-PCR identification and detection reagent for animal vesicular disease and preparation method and use thereof

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Embodiment Construction

[0033] 1. Construction of SVDV VP1, VSV N, FMDV 3D gene recombinant plasmid

[0034] According to Hua Qunyi et al. "Cloning and Sequence Analysis of Vesicular Stomatitis Virus Nucleoprotein Gene", Chinese Journal of Biological Products, Issue 1, 2004, pages 4 to 7, clone the N gene of Vesicular Stomatitis Virus And sequencing, construct SVDV VP1, VSV N, FMDV 3D gene gene recombination plasmid, named the recombinant plasmid pBAD-VP1, pBAD-VN5, pBAD-3D.

[0035] 2. Design primers

[0036]The present invention selects the conserved fragments of SVDV VP1, VSV N, FMDV 3D genes as the target, by comparing the DNA sequences of SVDV, VSV, FMDV genes reported in GenBank and the above 1 cloned and sequenced SVDVVP1, VSV N, FMDV 3D gene DNA Sequence homology analysis and comparison, respectively selected the conservative fragments of SVDV VP1, VSV N, FMDV 3D gene (126bp, 301bp and 189bp), using primer Express software and primer prere5.0 software, design primers, primer synthesis using β -Ac...

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Abstract

The related biological agent designed with target object as virus specific and conservative gene fragment of SVDV, VSV and FMDV, special the conservative fragment of VP1, N gene and 3D gene. Wherein, the RT-PCR detection agent includes three couple specific primers, and the amplification target fragment length is 126bp, 301bp and 189bp respectively. This invention can detect three viruses (SVDV, VSV and FMDV) in one reaction tube simultaneously.

Description

Technical field [0001] The present invention relates to an animal vesicular disease such as swine vesicular disease virus (SVDV), vesicular stomatitis virus (VSV), foot-and-mouth disease virus (FMDV) virus-specific and conservative gene fragments designed and synthesized biological preparations for the target, especially It is a reagent capable of simultaneously identifying and detecting three viruses of SVDV, VSV and FMDV, as well as the preparation method and application of this reagent. Background technique [0002] Swine vesicular disease (SVD), also known as swine vesicular disease, is an acute, febrile, and contact infectious disease caused by swine vesicular virus (SVDV). It is characterized by vesicular damage to the mouth and hoof. Its clinical symptoms are very similar to swine foot-and-mouth disease. It was first discovered in the pig herd of Lombardy, Italy in October 1966. Afterwards, the epidemic spread of swine vesicular disease occurred in many European countries,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 花群义周晓黎董俊杨云庆徐自忠肖荣海尹尚莲
Owner CHECKOUT & QUARANTINE TECH CENT YUNNAN ENTRY &EXIT CHECKOUT & QUARANTINE BUR
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