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PCR (Polymerase Chain Reaction) detection primer and method for H120 strain of avian infectious bronchitis virus

A bronchitis and H120 technology, applied in the field of PCR detection primers of avian infectious bronchitis virus H120 strains, can solve the problems of inconvenience in rapid detection of IBV, and achieve the effects of small fragments, time saving and high primer specificity

Inactive Publication Date: 2012-10-10
WENS FOOD GRP CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At this stage, the method of distinguishing vaccine strains from wild virus strains is through RT-PCR, and the amplified products are directly sequenced or sequenced after cloning, and then the results can be obtained after sequence analysis. It usually takes 3 to 7 days. For IBV The rapid detection of

Method used

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  • PCR (Polymerase Chain Reaction) detection primer and method for H120 strain of avian infectious bronchitis virus
  • PCR (Polymerase Chain Reaction) detection primer and method for H120 strain of avian infectious bronchitis virus
  • PCR (Polymerase Chain Reaction) detection primer and method for H120 strain of avian infectious bronchitis virus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1 Test material

[0022] 1.1 Main reagents and instruments

[0023] AxyPrep Body Fluid Viral DNA / RNA Mini Kit (Cat No.: AP-MN-BF-VNA-250), TaKaRa PrimeScript 0ne step RT-PCR Kit Ver.2 (TaKaRa Code: DRR055A), Thermo high-speed low-temperature centrifuge, Thermo PCR machine.

[0024] 1.2 Strains

[0025] IBV wild strains of each branch isolated clinically, Dahuanong Newcastle disease and infectious bronchitis dual live vaccine (H120 strain + Lasota strain; purchased from Guangdong Dahuanong Animal Health Products Co., Ltd.; production batch number: 201136), Kazakhstan Dahuanong Newcastle Disease and Infectious Bronchitis Dual Live Vaccine (trade name "Xinzhipic"; H120 strain + Lasota strain; purchased from Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences; production batch number 110514).

[0026] 1.3 Primer design

[0027] According to the sequence of IBV H120 strain (Genbank: FJ888351), a pair of primers were designed on the S1 gene an...

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Abstract

The invention discloses a PCR (Polymerase Chain Reaction) detection primer and method for H120 strain of avian infectious bronchitis virus, belonging to the technical field of detection of avian virus. The PCR detection primer comprises two pairs of detection primers, wherein the nucleotide sequences are respectively SEQIDNO: 1-2 and SEQIDNO: 3-4. By use of the two pairs of primers, RNA (Riboneucleic Acid) of a sample to be detected is used as a template so as to carry out RT-PCR amplification, and if products of two types such as 653bp and 230bp are amplified simultaneously, the containing of the H120 of the IBV in the sample to be detected can be judged. The PCR detection primer has the advantages that the specificity of the primer is high, the segments are small, the whole RT-PCR process can be finished in 2-2.5 hours, the H120 vaccine strain of the IBV can be quickly identified without the sequencing need of the sequencing company, so that the time is greatly saved for clinic detection.

Description

technical field [0001] The invention relates to the technical field of poultry virus detection, in particular to PCR detection primers and a detection method for avian infectious bronchitis virus H120 strain. Background technique [0002] Avian infectious bronchitis (IB) is an acute, highly contagious disease caused by avian infectious bronchitis virus (IBV). Under large-scale breeding conditions, the infection rate of avian infectious bronchitis is very high, the mortality rate of the disease is 10-30%, and the annual economic loss is more than one billion yuan. The H120 strain is the earliest attenuated vaccine used to prevent IB, and it is also one of the most widely used IBV attenuated vaccines. The genotyping belongs to the IBV branch V, and it has good safety and stable virulence. [0003] Since the avian infectious bronchitis vaccine commonly used in production is generally the new branch dual-linked attenuated live vaccine, the live virus of the vaccine virus exists...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 王峰
Owner WENS FOOD GRP CO LTD
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