Multiple PCR primer used for simultaneously detecting infectious Bovine Rhinotracheitis virus and akabane virus as well as its design method

A technology for rhinotracheitis virus and Akabane virus, which is applied in the field of multiplex PCR primers and their design, can solve problems such as shortening detection time and detection cost, and achieve the effects of protecting health, rapid development, strong specificity and high sensitivity

Inactive Publication Date: 2013-06-19
厦门佰能检验技术服务有限公司
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  • Abstract
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Problems solved by technology

The improved technology of multiplex PCR simplifies the detection process of food microorganisms, shortens the detection time, reduces the detecti

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  • Multiple PCR primer used for simultaneously detecting infectious Bovine Rhinotracheitis virus and akabane virus as well as its design method
  • Multiple PCR primer used for simultaneously detecting infectious Bovine Rhinotracheitis virus and akabane virus as well as its design method
  • Multiple PCR primer used for simultaneously detecting infectious Bovine Rhinotracheitis virus and akabane virus as well as its design method

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Embodiment Construction

[0036] The present invention will be further described below with reference to the drawings and specific embodiments, but the drawings and specific embodiments should not be construed as limiting the present invention.

[0037] The method for designing multiple PCR primers for simultaneous detection of bovine infectious rhinotracheitis virus and Akabane virus includes the following steps:

[0038] a) Use Primer Premier5.0 primer design software to design multiple primer combinations that simultaneously amplify bovine infectious rhinotracheitis virus and Akabane virus.

[0039] b) Screen the primers for PCR detection of bovine infectious rhinotracheitis virus in multiple primer combinations.

[0040] c) Screen the primers for PCR detection of Akabane virus in multiple primer combinations.

[0041] d) Screening multiplex PCR technology for simultaneous detection of primer combinations for bovine infectious rhinotracheitis virus and Akabane virus.

[0042] e) Determination of optimal reacti...

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Abstract

The invention relates to a multiple PCR primer used for simultaneously detecting infectious Bovine Rhinotracheitis virus and akabane virus as well as its design method, and relates to detection of infectious Bovine Rhinotracheitis virus and akabane virus. The size of a PCR amplification fragment corresponded to an infectious Bovine Rhinotracheitis virus primer in the multiple PCR primer is 311bp, the size of the PCR amplification fragment corresponded to akabane virus id 392bp. The method comprises the following steps: 1) designing multiple PCR primer combination; 2) screening the primer in the primer combination, keeping the primer which can not synthesize a primer dimer; 3) determining the competition advantage and disadvantage states of the kept primer, comparing GC% and base number in the kept primer, selecting the kept primer with high GC content and determining as the primer with excellent competition state, performing a step 5); otherwise, performing a step 4); 4) selecting the primer with poor competition state again, wherein the concrete step repeats the step 2) and determining the kept primer according to the step 3) again; and 5) performing amplification on the primer with the excellent competition state.

Description

Technical field [0001] The invention relates to the detection of bovine infectious rhinotracheitis virus and Akabane virus, in particular to a multiple PCR primer for simultaneous detection of bovine infectious rhinotracheitis virus and Akabane virus and a design method thereof. Background technique [0002] Infectious bovine rhinotracheitis (IBR) is an acute febrile respiratory infection of cattle. It was first discovered in beef cattle herds in the western United States in 1950, and later appeared in dairy herds. The virus was isolated in 1956 and was identified as a bovine herpes virus. It is clinically prone to bovine epidemic fever, bovine flu, malignant catarrhal fever and bovine mucosal disease Syndrome is confused. The clinical features of IBR are fever, upper respiratory tract mucosal inflammation, edema, bleeding, and even necrotic spots. Infectious vaginal vaginitis (IVP) occurs after the cow is infected. It can also induce bovine infectious balanitis, conjunctiviti...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12N15/10C12R1/93
Inventor 孔繁德徐淑菲陈圣军张吉红唐泰山陈信忠龚艳清郭书林杨俊萍
Owner 厦门佰能检验技术服务有限公司
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