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

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...

AI Technical Summary

Problems solved by technology

The improved technology of multiplex PCR simplifies the detection process of food microorganisms, shortens the detection time, reduces the detection cost and improves the sensitivity of food microorganism detection; however, there are some problems in the improved technology of multiplex PCR

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