Multiplex PCR kit for simultaneously detecting four viruses carried by ruminants

A technology for ruminants and kits, which is applied in the field of multiplex PCR kits and multiplex PCR detection kits, can solve the problems of time-consuming and laborious, easy to confuse clinical diagnosis, difficult to distinguish clinical diagnosis, etc., and achieve high detection sensitivity and good reproducible effect

Active Publication Date: 2015-03-25
艾军 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After being infected with one of these four viruses, the sick animals all showed symptoms such as depression, body temperature, mucosal edema and erosion clinically, which were not easy to distinguish in clinical diagnosis
For ruminant clinical mixed infection diseases - bluetongue, foot-and-mouth disease, vesicular stomatitis, Peste des petits ruminants, clinical diagnosis is easy to confuse, traditional virology test is time-consuming and labor-intensive, ordinary PCR detection requires different reaction systems, time-consuming Due to the laborious characteristics, strengthening the research and development of rapid detection reagents is the demand for epidemic detection

Method used

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  • Multiplex PCR kit for simultaneously detecting four viruses carried by ruminants
  • Multiplex PCR kit for simultaneously detecting four viruses carried by ruminants
  • Multiplex PCR kit for simultaneously detecting four viruses carried by ruminants

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Experimental program
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Embodiment

[0077] 1 Materials and methods

[0078] 1.1 Positive\negative control

[0079] Positive controls: plasmids carrying bluetongue virus NS3 gene, plasmids carrying foot-and-mouth disease virus 3D gene, plasmids carrying PPR vaccine strain N gene, and plasmid mixtures carrying vesicular stomatitis virus N gene were provided by Yunnan Entry-Exit Inspection and Quarantine Bureau The Animal Inspection Laboratory and the Animal Inspection Laboratory of Chongqing Entry-Exit Inspection and Quarantine Bureau kept it and donated it to the Animal Inspection Laboratory of Shenzhen Entry-Exit Inspection and Quarantine Bureau; the public can obtain it from any of the above units, or entrust Dalian Baobio Biotechnology Co., Ltd. Or other biological limited synthesis.

[0080] Negative control: deionized water without PPRV, BTV, VSV and FMDV.

[0081] 1.2 Primer solution

[0082] The primers in Table 1 were synthesized by Dalian Bao Biological Co., Ltd. Different primers were configured at 20 ...

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Abstract

The invention discloses a multiplex PCR kit for simultaneously detecting four viruses carried by ruminants. The kit comprises four pairs of specific primers. The experiment proves that bluetongue, foot-and-mouth disease, peste des petits ruminants and vesicular stomatitis virus nucleic acid in clinical samples of ruminants can be simultaneously detected. The kit has the characteristics of high sensitivity, high specificity and simplicity in operation, the detection time can be saved, and the condition that DNA contamination occurs in detection is reduced.

Description

technical field [0001] The invention relates to a multiplex PCR kit for simultaneously detecting four viruses carried by ruminants; in particular, it belongs to a multiplex PCR detection reagent for simultaneously detecting ruminant bluetongue, foot-and-mouth disease, Peste des Petits Ruminants and vesicular stomatitis viruses The box belongs to the field of virus detection. Background technique [0002] Multiplex PCR (multiplex PCR) is an improvement on the basis of ordinary PCR. Multiple pairs of specific primers are added to a PCR reaction system to amplify multiple target fragments for multiple DNA templates or different regions of the same template. Multiplex PCR is mainly used for the simultaneous detection or identification of multiple pathogenic microorganisms and the typing and identification of pathogenic microorganisms, certain genetic diseases and cancer genes. The characteristics of multiplex PCR are: 1. High efficiency, multiple pathogenic microorganisms can b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/701C12Q2600/16
Inventor 艾军秦敏祝贺聂福平蒋宏伟韩佃刚张滨杨云庆叶玲玲周晓黎花群义李应国董俊
Owner 艾军
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